Module 1 Unit 5 Bio Chemistry

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Module 1 Unit 5 Bio Chemistry
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Society of Cosmetic Scientists Distance Learning
Unit 5
Prepared by David Peers with Grace Abamba, Angela Beattie, David Benzies, Rachel Benzies and Brian Knight
DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5
CONTENTS 1 Bio chemical Materials Carbohydrates 1.1 Introduction 1.2 Basic Carbohydrates Activity 1 Structure of simple carbohydrates 1.3 Bigger Molecules Activity 2 Hemose Sugars Activity 3 Ring Structures of Sugars 1.4 More Complex Carbohydrates Activity 4 More Complex Sugars 1.5 Polysaccharides 1.6 More Complex Polysaccharides 1.7 Uses of Carbohydrates i) Energy Supply ii) Structural iii) Water Trapping 1.8 Summary Checklist 2 Lipids 2.1 Introduction 2.2 Biological fats and oils 2.3 The nature of fatty acids 2.4 The implications of unsaturation Activity 5 Double Bonds in Hydrocarbon Chains 2.5 Other classes of Lipids Activity 6 Comparison of Lipid Structures 2.6 Uses of Lipids 2.7 Summary Checklist Appendix 1 3 Proteins 3.1 Introduction 3.2 Amino Acids Activity 7 Optical Isomers of Amino Acids Activity 8 Inuence of Amino Acid Sidechains 3.3 Peptides Activity 9 Formation of a Tripetude 3.4 Proteins 3.5 Protein Structure 3.6 Tertiary Structure 3.7 Forces Invloved in Maintaining Tertiary Structure 3.8 Quaternary Structure Activity 10 Levels of Protein Structure 3.9 Summary Checklist 4 4 4 5 6 7 8 9 9 10 12 12 12 12 13 13 14 15 15 15 15 17 17 19 20 21 22 22 23 25 25 25 25 28 29 30 32 32 35 36 38 40 40 41
BIO CHEMISTRY
4 Enzymes 4.1 Introduction 4.2 Function of Enzymes 4.3 Enzyme Classication Activity 11 Classication of Enzymes 4.4 Enzyme Structure 4.5 The Enzyme Substrate Complex Activity 12 Enzyme Structure and Mechanism 4.6 Enzyme Inhibition 1) Irreversible Inhibitors 2) Reversable Inhibitors Activity 13 Effect of Different Inhibitors 4.7 Summary Checklist 5 Metabolic Pathways 5.1 Introduction 6 Genetics and Cell Replication 6.1 Introduction 6.2 The Structure of DNA Activity 14 Base Structure of DNA Activity 15 The Principle of Complementary Base Pairs in DNA Checklist 6.3 Replication of DNA Activity 16 The Semi-Conservative Process of DNA Replication 6.4 DNA Transcription Activity 17 Processes Involving DNA and RNA 6.5 Summary Checklist Appendix 2 Appendix 3 Appendix 4
42 42 42 43 43 44 44 45 47 47 47 47 49 49 50 50 52 52 52 54 56 57 57 58 58 61 61 62 63 64 65
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1 BIO CHEMICAL MATERIALS CARBOHYDRATES 1.1 Introduction Bio chemistry involves thousands of different chemical compounds, which can be sorted into a number of classes. Their application to Cosmetic Science is that these are the substances which make up human bodies. These are the substances to which cosmetic products are applied. Many of them are substances from which cosmetic products are manufactured, as will be explained during the Course. As this is an introductory unit, the basic nature of these substances will be treated in a more biological context, that of food. This treatment explains the ways in which the underlying arrangements of chemical structure, often simply physical size and shape, underlie their properties in real, everyday practice. In the next three sections, we will examine the structures and properties of three major classes of bio chemical molecules, carbohydrates, lipids and proteins. Make sure you have available your molecular model kit, provided in the Practical Activity Kit. 1.2 Basic carbohydrates As the rst group of bio chemical substances to be considered, these will be discussed in some depth. Many are used in the cosmetics industry, for example, modied celluloses are common thickening agents. The basic principles established here, that underlying structures and patterns of bonding inuence the properties of real substances, will then be applied to other materials, such as oils and proteins. Carbohydrates contain the chemical elements carbon, hydrogen and oxygen. In the overall molecular formula there are always twice the number of hydrogen atoms as there are oxygen atoms. For example, the carbohydrate substance which is most familiar in everyday life is probably table sugar sucrose. It has the overall formula C12H22O11. Actually, in general bio chemical terms, sucrose is something of an irrelevance. It is only important to sugar beet plants, sugar cane plants and humans. To a biologist, the word sugar tends to mean glucose, C6H12O6, and this is the example that is given rst in most textbooks. Why do you think that, other than perversity, I chose sucrose? One reason is its familiarity. The average consumption in Britain is currently 110 g/day per head, giving a yearly total for the UK of 2.2 million tonnes. (Source: Sugar Bureau.) And, if you think you dont eat that much, there are 9 teaspoons (45 grams) in a can of soft drink. If you are not eating this much, somebody is getting more than their fair share. And this is simply rened sucrose. There are many other sources of sugars. More pertinent, in my experience, is the fact that people who meet the formula for glucose rst develop the lasting xation that the number of carbon atoms in the molecule of carbohydrate must equal the number of oxygen atoms. Look at the formula for sucrose and count the different atoms: C12H22O11. From this you can see that any carbohydrate will t a general formula, CxH2yOy, where x and y can be any number from 3 to several thousands. (Put this slightly differently, Cx(H2O)y, and you have carbon hydrate.) The smallest, simplest sugars have 3 carbon atoms and are therefore called trioses. We shall explore the possibilities of various arrangements of their components, since the principles also apply to more complicated examples. From our denition of the proportions of the chemical elements found in carbohydrates, each of the three carbon atoms must be attached to the elements of water: H- and -OH.
BIO CHEMISTRY
ACTIVITY 1 Structure of simple carbohydrates (Allow about 15 minutes) I am going to ask you to assemble molecular models, in order to explore the various arrangements that are possible using the elements of a simple carbohydrate and which qualify as carbohydrates. In doing so, I hope you will begin to understand how the molecular structures and their physical shapes inuence their properties. The principles you derive from assembling these models will be extremely useful later in the course and in your future work. 1) Assemble a chain of three carbons (black). Add to each the elements of water, hydrogen (white) and hydroxyl (white for hydrogen and red for oxygen). If you were to write a structural formula for the molecule that you have constructed it would look like this:
H C O H H C O H H C O H
Unfortunately, as a molecular structure, this will not do because we have two spare bonds unlled. The solution would seem to be to simply plug them with hydrogen atoms. This still will not do. By adding more hydrogens, we have deviated from our basic criteria. (However, do not dismiss this new molecule. It represents a real substance, which we shall meet again. It is glycerol.) 2) Rearrange our rst molecular structure, as shown above, containing the atoms C3H6O3 to produce viable molecules, with each element showing its correct valency (carbon 4, hydrogen l and oxygen 2). Those two spare bonds must be accounted for, in some way. Do this now and draw structural formulae for your suggestions below. If this causes you any hesitation, then you really should go back and work through Unit 1, Basic Chemistry. (Production of double bonds will involve bending the bonds in your molecular model kit.)
BIO CHEMISTRY
There are several possibilities. I have reproduced two in gure 1.

Carbonyl group C O H C O H C OH H C OH H H Aldehyde group H H C OH C O C OH H Aldose sugar Ketose sugar Ketone group
Figure 1. Possible arrangements of 3C: 6H: 3O
The rst is known as an aldose sugar., because its carbonyl group (-C=O) is on the end of the molecule as part of an aldehyde group.
H C O
The second is known as a ketose sugar, because -C=O is in the middle of the molecule as part of a ketone group.
C O
Both arrangements are trioses. They represent the simplest examples of two families of sugars. They are isomers of each other. If you are not convinced that this sleight of hand represents different chemical substances, think of typical, day-today examples. Aldehydes provide many of the off-avours and smells in old foods think of the smell of over-ripe apples. The best known ketone is acetone, nail varnish remover. These are obviously different substances. These two triose sugars are not met in everyday life and do not have common names. However, they are important in the pathways of chemical reactions which make up our metabolism and we shall meet them again in later sections. Most sugars also display other forms of isometry, geometric and optical (or stereo) isometry. We dont need to worry about this at the moment. We can now represent the chemical formula for a simple sugar as a chain of carbon atoms, most carrying the elements of water. One carbon atom will have a carbonyl group (-C=O), either on the end of the molecule (aldehyde) or in the middle (ketone), giving rise to two families of sugars, the aldoses and the ketoses. 1.3 Bigger molecules If you go back to your original molecular model structure, you can see that we can still meet our criteria for a carbohydrate even if we insert more and more sets of
H C OH
into our theoretical structure. This would give us larger molecules of sugars, tetroses (with four carbon atoms), pentoses (with ve) and so on. The important ones, for now, are the hexoses (with six carbon atoms). In these larger molecules, even more variations in arrangement are possible. On the following page, in gure 2, I have drawn the two most important versions of a hexose.
BIO CHEMISTRY
CHO Unspecific representation (no information on structure) HO C H HO C H HO C H H C OH CH 2OH
Glucose: an aldo-hexose Carbon atom number H C O H C OH HO C H H C OH H C OH H C OH H 1 2 3 4 5 6
Fructose: a keto-hexose
H H C OH C O HO C H H C OH H C OH H C OH H
Figure 2. Structure of hexose sugars
The numbering system for the carbon atoms is conventional, counting from the end nearest to the carbon atom which carries the most reactive functional group, in this case the carbonyl -C=O, towards the top of the formula in gure 9; carbon atoms l and 2 respectively.
ACTIVITY 2 Hexose sugars (Allow about 3 minutes) a) Add up the numbers of different atoms in both glucose and fructose and write them below.
Number of atoms present Carbon Glucose Fructose Hydrogen Oxygen
Both are C6H12O. They are isomers of each other. b) Assemble a molecular model of glucose. Make sure that you have all the groups in the correct orientation. In the written formula, the hydroxyl group on carbon number 3 points the opposite way. Make sure that this is reected in your model. This is geometric isometry.
BIO CHEMISTRY
These open chain formulae for sugars have been useful in showing us that carbohydrate molecules have basic similarities that can be adapted to give a variety of substances. They are called Fischer structures, after the nineteenth-century chemist who was a pioneer in carbohydrate chemistry. Look at your molecular model of glucose, the sugar! You can see that the at, unexciting depiction on paper represents a real collection of material substance. It would be long and oppy. In a solution, buffeted by surrounding water molecules, it would twist about. Chemical bonds are of precise length and angle. In practice, the aldehyde group, on carbon number l, would come close enough to the hydroxyl on the fth carbon atom in the chain to interact with it. A ring structure is formed (the Fischer formulae have been tightened up):
CHO HCOH HOCH HCOH HCOH H2COH
HCOH HCOH HOCH HCOH HC H2COH O
A better way of showing the ring is shown in gure 3. (The numbers reect the carbon atoms, counting from what would have been carbon number l in the open chain structure, that is the carbon atom carrying the most chemically reactive group, the aldehyde.)
CH2OH O H H
2C
H
4C
C
5
H
1C
HO
OH
3C
OH
H
Figure 3. Glucose as a ring structure
OH
ACTIVITY 3 Ring structures of sugars (Allow about 3 minutes) Adapt your molecular model into the ring format shown in gure 10. Again, make sure that you have the correct orientation for all groups.
In forming this ring structure and handling your completed model, you will have a better sense of a three-dimensional substance. It has been estimated that more than 99% of glucose molecules in solution are in such ring forms.
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What would a fructose molecule look like in a ring form? Once again, the most reactive group is the carbonyl, in this case being a ketone on carbon number 2. Once again, it reacts with carbon number 5, giving, in this case a ve-membered ring, with two offshoots, as shown in gure 4.
6CH2OH
O
5C
OH
2C
H
4C
HO
3C 1CH OH 2
OH
Figure 4. Fructose as a ring structure
1.4 More complex carbohydrates
ACTIVITY 4 More complex sugars (Allow about 5 minutes) Earlier, I said that the basic principles for production of a molecule of carbohydrate could be extended to produce larger and more complex molecules. Now, the ring structures, produced in Activity 9, will be combined to form even more complex sugars. Take your ring structure for a molecule of glucose, construct another copy and align them as shown below:
CH2OH O H H
2C
CH2OH O H H
2C
H
4C
C
5
H
1C
H
4C
C
5
H
1C
HO
OH
3C
OH
HO
OH
3C
OH
OH
OH
If we remove the elements of water, shown shaded above, joining the two molecules, we have a particular class of chemical reaction. (It is called condensation. The reverse, splitting a larger molecule with insertion of water, is called hydrolysis.) This gives us the condensed sugar shown in gure 5.
BIO CHEMISTRY

CH2OH H HO O H OH H H OH H O H CH2OH O H OH H H OH H OH
Figure 5. Condensed sugars
You have assembled an example of a disaccharide, formed by joining two monosaccharides. (It is worth emphasising that saccharins (articial sweeteners) have no chemical relationship to sugars.) A number of disaccharides are signicant in day-to-day life. The one we have constructed by combining two units of glucose is called maltose but others are possible as illustrated below: GLUCOSE + GLUCOSE GLUCOSE + FRUCOSE GLUCOSE + GALACTOSE gives MALTOSE gives SUCROSE gives LACTOSE (milk sugar)
Look again at your model or the formula. We still have an hydroxyl group attached to a carbon number l on one end of the molecule and an hydroxyl group attached to a carbon number 4 at the other end of the molecule. If we line up yet another glucose molecule, we can carry out another condensation, to give a trisaccharide. This can be repeated many times to give a polysaccharide. 1.5 Polysaccharides There are several variations we can create simply by combining glucose molecules in differing ways. The one we have just created, a long wavy chain made up of 200- 300 glucose subunits joined by 1-4 bonds, is called amylose. There are other possibilities. Look back at a glucose molecule. There are 5 hydroxyl groups sticking out from the ring. Those on carbons number 1 and 4 are the most likely to react together, because they are sticking out at the ends. As the molecules jostle around in solution in water, they are simply the hydroxyl groups which are most likely to bang into each other, giving the possibility of a chemical reaction. However, other groups can also react. One possibility is shown in gure 6.
HO H OH H H CH 2OH O H HO H HO CH 2OH H HO O H OH H H OH H OH H disaccharide O HO HO H HO HO H H O H CH 2 H H OH H O OH
CH 2OH
Figure 6. Alternative possibility for linking glucose molecules
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There is the possibility of a 1-6 bond. This produces a molecule which consists of chains of glucose units, joined by 1-4 bonds, as in amylose above. About every twenty ve glucose units, a 1-6 bond starts a branch, giving the sort of pattern illustrated in gure 7:
about 25 units
Figure 7. Branching arrangement of polysaccharide chains
This molecule is called amylopectin. Amylose and amylopectin are not household names, but starch is a mixture of the two. Starch from the wheat plant (our) consists of about one third amylose and two thirds amylopectin. We have seen that different shapes and alignments enable molecules to interact in various ways. Amylose is a loose, oppy chain, so a starch with a higher proportion of this would be looser; potato starch is an example. Amylopectin is a more compact molecule, more of this gives the waxy starches, such as from peas. It is easy to start thinking of these molecules as abstractions. Notice again how their shapes and arrangements affect their reality. The cosmetics industry must pay attention to ways of affecting consistency and texture. Plants produce starches. Animals such as ourselves produce glycogen. This is similar in principle to amylopectin, but is more highly branched, about every 12 glucose subunits. There is one nal variation on polysaccharide formulae based solely on glucose units. When we converted our open chain formula into the ring form, we bent it one way. It could also have bent the other way, as shown in gure 8.
6
CH 2OH OH H C1 C4
CH2 OH OH O C1
5C
5C
C4
3C
C2
3C
C2
CH 2OH
5
CH 2OH
5
O
1 2 1 4
OH
1 1
OH
3
-glucose
Figure 8. Alternative ring structures for glucose
-glucose
Chains of glucose units formed from 1-4 bonds between molecules of betaglucose are the basis of cellulose. The practical implications will be dealt with later.
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1.6 More complex polysaccharides The examples we have met so far are based solely on glucose units, combined in various ways. Yet, we managed to produce four distinct substances. Think of the range of possibilities if we could use two, three, or even more types of subunits in the same polysaccharide. This gives rise to a wide range of complex carbohydrates, such as the pectins and hemicelluloses. In combination with other bio chemical materials we obtain the glycoproteins, mucopolysaccharides and so on. Some of these will be mentioned under Uses of carbohydrates (see below) but the full range is outside the scope of this unit. 1.7 Uses of carbohydrates i) Energy supply Glucose is the basis of biological energy. It is produced by plants by the process of photosynthesis, as summarised below. Carbon dioxide CO2 + water + energy (sunlight) + H2O + energy = glucose = C6H12O6 + oxygen + O2
Plants then convert the glucose to starch for storage. At some future time, when the plant needs a supply of energy for respiration, it must break down the starch again to give free glucose. But why doesnt it simply store the glucose to begin with? In my experience, most people give an immediate answer based on the glucose somehow leaking out of the cell. In fact, there are problems getting sugars through cell membranes, which are made of fatty materials. Think back to your experiment with the egg and the sugar solution. The naked cell membrane was surrounded by a concentrated solution of sugar. The molecules of sugar could not pass through the fatty membrane, so we had osmosis. In the present case, the cells of the plant would attempt to build up a concentrated solution (of glucose), surrounded by a cell membrane. Water would be drawn in by osmosis. The cells would swell. By combining the glucose molecules and storing granules of starch, which are, to all practical purposes, insoluble in these conditions the cells avoid this difculty. Similarly, we use glucose as a simple, soluble means of transporting carbohydrate. We store insoluble polysaccharide (glycogen). ii) Structural Cellulose is probably the most widespread bio chemical molecule, strengthening the cell walls of plants and bacteria. We have seen that its beta 1-4 bonds are a different shape from alpha 1-4 bonds, such as in starch. (No animal can make an enzyme which can break down such bonds. We have amylases, which break down alpha 1-4 bonds, for example, in saliva. We cannot digest beta l-4 bonds. Animals which eat exclusively plant-based foods rely on specially modied guts which harbour micro-organisms, for example in the pouches on the stomachs of cows or in the appendix of rabbits. These microbes can produce cellulase enzymes.) Have another look at your glucose model and note that the hydroxyl groups stick out. In the chains of cellulose molecules, these can interact and form hydrogen bonds, which bind the chains together, so the cellulose chains are locked together into rigid laments, giving support to the plant cell walls and, incidentally, making it even harder for animals to digest. Cellulose makes up a large proportion of bre in the diet. Hydrogen bonds are weak. However, imagine, that instead of a molecule being made up solely of glucose subunits, every half dozen or so along the chain we have some other chemical subunit. Where two chains lie close, these can form strong chemical bonds, and so on to form a complex interlocking three-dimensional network which could be immensely strong. This is the basis of complex structural carbohydrates in both animals and plants. Plants growing more than a couple of feet in height must develop more woody support; think of the difference between a small and a tall nettle plant. The Californian Redwood tree can grow to over 200 feet. Ebony is harder than mild steel. The animal equivalent is chitin in the external skeletons of arthropods. This allows the South American Goliath Beetle to grow to 9 inches. Any larger than this and the weight of the skeleton becomes overwhelming. In water, which provides support, the Japanese Spider Crab can reach 12 feet across the legs, all supported by complex interlocking carbohydrates.
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Hydrogen bonds are weak. However, imagine, that instead of a molecule being made up solely of glucose subunits, every half dozen or so along the chain we have some other chemical subunit. Where two chains lie close, these can form strong chemical bonds, and so on to form a complex interlocking three-dimensional network which could be immensely strong. This is the basis of complex structural carbohydrates in both animals and plants. Plants growing more than a couple of feet in height must develop more woody support; think of the difference between a small and a tall nettle plant. The Californian Redwood tree can grow to over 200 feet. Ebony is harder than mild steel. The animal equivalent is chitin in the externa skeletons of arthropods. This allows the South American Goliath Beetle to grow to 9 inches. Any larger than this and the weight of the skeleton becomes overwhelming. In water, which provides support, the Japanese Spider Crab can reach 12 feet across the legs, all supported by complex interlocking carbohydrates. Complex, interlocking carbohydrates also have a human structural role, for example in the threedimensional, water-trapping meshworks (below) which form the matrix or ground substance of connective tissues. An example sometimes used as a component in cosmetics is hyaluronic acid in the dermis of the skin. We have seen that chains made up from simple sub-units can interact with each other, to form complex, three-dimensional structures. If the basic chains also include different sub-groups, which can form strong chemical bonds with each other, then these three-dimensional structures can be very strong, providing support for living organisms. iii) Water trapping Even starch, a comparatively simple polysaccharide, can trap water to form pastes, as in making custard or white sauce. This brings together many of the points we have been exploring about dayto-day properties of materials being dependent on their shapes and interrelationships. In heating and stirring, the our is broken up, and starch chains spread through the liquid. As it cools, the chains form hydrogen bonds in a three-dimensional network. This traps the liquid into a gel. Once again, more complex molecules can extend these possibilities. Imagine a towel. Think of the difference between a towel made from a smooth bre and one made from a uffy bre. The latter soaks up more water. Extend this image to separate molecules. Starch is largely simple chains. A more highly branched molecule (uffy) can trap more water. An example is the dermis of the skin which has a matrix containing complex polysaccharides, such as hyaluronic acid. Retention of this water is important for the integrity of the skin. It reduces with age. Glycogen, in the liver, has large amounts of associated water. It is loss of this, long before any signicant loss of fat, which accounts for impressive weight losses in the rst week of slimming diets. Examples of even more impressive water-trapping properties include extracts from seaweeds (agar-agar is well known in microbiological plates and alginates are used both in the food and cosmetics industries) and mosses (carrageenates). The record achievement, in terms of water trapping, is probably from the latter, 1 gram of which can lock 11 litres of water into a gel. 1.8 Summary Carbohydrates are a widespread group of substances which can be used to illustrate a number of basic principles of bio chemical organisation. They can be divided into two broad classes. Comparatively small molecules (sugars) show that a range of substances can be produced by different arrangements of similar chemical units. These can, in turn, be combined into large, complex molecules (polysaccharides). The properties of these depend to a large extent on simple principles of physical shape and interaction. Important bio chemical functions of carbohydrates which emerge from these properties include energy supply and storage, structural support and water retention.
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CHECKLIST At the end of this section you should be able to: draw a structure for a simple carbohydrate such as glucose use molecular models to distinguish between isomers of carbohydrates, such as aldoses and ketoses describe and illustrate how, through the process of condensation, molecules of simple sugars can combine to form more complex molecules describe and illustrate how, by extension of the same principles, a variety of large, complex molecules of polysaccharide can be produced explain how the properties of these substances, in realistic day-to-day practice, are based on their shapes and interconnections, with reference to energy supply, energy storage, water trapping and rigid, structural molecules.
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2 LIPIDS 2.1 Introduction Lipids are not so clear cut as a category of molecules as carbohydrates. They include the oily, greasy and waxy substances which are part of living processes. The cosmetics industry handles many such materials: some of these are biological lipids, some others are synthetic alternatives. Once again, as with many organic molecules, they consist of carbon, hydrogen and oxygen, but there are no clear proportions of these, as there are with carbohydrates. However, in general, there is much less oxygen in the molecules. The implications of this will be discussed later. Molecules of lipid contain high proportions of hydrogen and carbon. This means that, like the mineral hydrocarbons, they are relatively non-polar/hydrophobic; that is they will not mix well with water. In contrast, they will dissolve in organic solvents such as ether and chloroform. There are four main classes of lipids which are important in the context of the cosmetics industry: fats and oils; waxes; phospholipids and steroids. As with carbohydrates, I will consider the nature of the rst in some detail to establish basic principles. These can then be applied to the other classes. 2.2 Biological fats and oils These are technically called triacylglycerols. They were traditionally known as triglycerides or neutral lipids. They are the fats and oils found in biology and must be distinguished from the mineral oils. Other units, in particular, Unit 7 Oils, Fats and Waxes, will make this distinction apparent and apply the principles explored here to their applications in cosmetic products. Triacylglycerols are, chemically, esters between glycerol and long chain fatty acids. You will remember that an ester is a compound between an alcohol and a carboxylic acid. (If you are uncertain about any of these points, you should go back to Unit 1 Basic Chemistry.) We met glycerol (in everyday language, glycerine,) in the previous section when developing our understanding of the nature of simple carbohydrates. It has three carbon atoms, each carrying a hydroxyl (alcohol) group, as shown below in gure 9.
H2 C H C H2 C
Figure 9 The structure of glycerol
OH OH OH
There are 3 hydroxyl groups. It can therefore combine with three acid groups (hence, tri-acylglycerols). 2.3 The nature of fatty acids From your knowledge of basic organic chemistry, you will remember that the simplest homologous series was the hydrocarbons. These are basically inert, unreactive molecules. Fatty acids are composed of hydrocarbon chains with carboxylic acid end groups. They remain dominated by their long hydrocarbon chains and retain large amounts of such inertness. Their predominant features are that they are highly reduced (consisting of little but carbon and hydrogen) and non-polar. As with the mineral hydrocarbons, molecules of long chain fatty acids can contain the theoretical maximum number of hydrogen atoms (saturated) or can have double bonds between some pairs of carbon atoms, reducing the number of hydrogen atoms (unsaturated). A few fatty acids are much more common than others in bio chemical fats. Most of these are also familiar in cosmetic products. Some examples are listed in table 1.
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Table 1. Examples of common fatty acids Name Stearic acid Palmitic acid Oleic acid Linoleic acid Linolenic acid Arachidonic acid Degree of saturation Saturated Saturated Unsaturated (monounsaturated) Unsaturated (polyunsaturated) Unsaturated (polyunsaturated) Unsaturated (polyunsaturated) Molecular composition C18.0* C16.0 C18.1 C18.2 C18.3 C20.4
(* the first number denotes the number of carbon atoms in the chain; the second number denotes the number of double bonds in the chain)
Animals such as ourselves cannot produce these poly-unsaturated fatty acids and must obtain them from their diet. They are therefore known as essential fatty acids. We will discuss this further under Uses. A molecule of triacylglycerol will have a formula something like that shown below in gure 10, depending on which fatty acids are involved.
O H2C O O HC O O H2C O C C C CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH2 CH3
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2 CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2 CH3
CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2 CH2
CH2
CH2
CH2
CH2
CH2
CH2
CH2 CH3
CH2
CH2
CH2
CH2
CH2
CH2
CH2
Figure 10. Structure of a triacylglycerol
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2.4 The implications of unsaturation
ACTIVITY 5 Double bonds in hydrocarbon chains (Allow about 6 minutes) The presence of double bonds has important effects on the shapes of hydrocarbon chains, which can be demonstrated using a molecular model. Use your molecular model kit to construct a length of hydrocarbon chain. Join eight carbon atoms (black) into a chain. Fill the remaining bonds with hydrogen atoms. As you handle your model, you will see that it is long and exible. (Remember that the hydrocarbon chains in fatty acids are twice this length!) Remove one hydrogen atom from each of two adjacent carbon atoms, somewhere in the middle. It is now unsaturated. Construct a double bond between these two carbon atoms. (This will involve bending the green bonds from your model kit.) Handle your model and observe its shape around the double bond. Detach one end of the double bond and exchange it with the other hydrogen atom on that carbon. Handle and observe the difference. Sketch your two molecular models.
1.
2.
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A chain of carbon atoms joined by single bonds (saturated) will be exible. A double bond adds a point of rigidity. In fact, double bonds can be two different shapes. The snaky, twisting hydrocarbon chain can approach the double bond from one side and the continuation after the bond come out from the other side. This is called trans (across). Alternatively, the continuation can come out from the same side of the double bond. This is called cis. These are illustrated in gure 11 below and should resemble your sketch.
H C H C C C H H
saturated
trans
saturated
cis
saturated
Figure 11. The shapes of double bonds in hydrocarbon chains
An obvious question is, does this matter? Think about this in terms of the physical shapes of the molecules, the ways in which they can interact and how these underlie the properties of the real substance. Think about the difference between trying to store string compared with something with rigid bends, say, wire coat hangers. The string can be packed up tightly. Coat hangers have a perverse ability to tangle. Bits stick out. Look at gure 12, below.
Figure 12 cis unsaturated hydrocarbon chains preventing close packing
In gure 12 you can see that cis unsaturated fatty acids cannot pack closely together. So, if our lipid material has a high proportion of fatty acids containing cis double bonds, it will be more uid or oily. If it has more saturated fatty acids, packed together, it will be denser, a fat. A dayto-day illustration would be the difference between lard and corn oil. Animals, like us, tend to produce mainly saturated fatty acids, giving rise to solid fats. Plants produce more unsaturated fatty acids and therefore oils. Most naturally occurring unsaturated fatty acids have cis double bonds. Food processing produces trans double bonds. The health implications of this are controversial. We can manipulate these properties industrially. For example, plant oils can be hydrogenated, that is, their double bonds can be made saturated, to give solid margarines. The same principles are applied to the lipid materials used in cosmetic products, giving a way to manipulate consistency and texture.
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2.5 Other classes of lipids The biological waxes are similar in principle to the triacylglycerols, that is, they are esters of alcohols with fatty acids. However, they contain alcohols other than glycerol. I must emphasise again that, here, we are discussing waxes produced by living things, such as beeswax, which is used in cosmetic products. The term wax is also used to mean, like oil, a range of mineral substances, such as candle wax. It is also used to mean a wide variety of synthetic substances. Some of these are not organic (based on carbon), for example, silicone waxes, such as furniture polish. All these categories are used in cosmetic products. The phospholipids are the major lipid components of cell membranes. As the name suggests, they contain phosphate groups, as well as glycerol and fatty acids. The combination of this phosphate, together with other chemical groups, makes part of the molecule more polar (able to mix with water) than the other categories of lipids. This is illustrated in gure 13.
O H2 C O O HC CH 3 CH 3 N+ CH 3 CH 2 CH2 O O P OO CH2 O C C CH 2 CH 2 CH2 CH2 CH 2 CH2 CH 2 CH2 CH 2 CH2 CH 2 CH3
CH 2
CH 2 CH2
CH2
CH 2 CH 2
CH 2
CH 2 CH 2
CH2
CH 2 CH2
CH 2 CH2
CH2
CH 2 CH3
CH 2
CH 2
CH2
CH 2
CH 2
polar/hydrophilic will mix with water
nonpolar/hydrophobic will not mix with water
Figure 13. Structure of a phospholipid
These intermediate properties make them excellent emulsifying agents. The best known group of phospholipids, the lecithins, are widely used in food processing. Their properties are used by the cosmetics industry in the production of liposomes. The cosmetics industry has a range of other substances which can be used to stabilise their oil and water systems. Some of these would not be palatable. (This topic is further discussed later in the course.) The nal class of lipids is the steroids. A typical structure is shown in gure 14.
CH3 CH3 CH2 CH2 CH2 CH2 CH HO CH3 C C CH2 CH CH CH CH2 C CH CH CH
CH2 CH2 CH2 CH2 CH CH3 CH3
CH2
Figure 14. Structure of a typical steroid
The basic steroid is cholesterol, illustrated in gure 14. It has a poor image, due to its association with heart disease. However, it is an essential bio chemical, produced in the liver, as a component of cell membranes and as the basic material for producing the steroid hormones, such as oestrogens and cortisones.
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ACTIVITY 6 Comparison of lipid structures (Allow about 5 minutes) We have met a variety of classes of substances which fall into the general category of lipids. A comparison will reinforce the theme of the relationship between structure and properties. Compare and contrast the major classes of lipids: triacylglycerols (gure 10); phospholipids (gure 13) and steroids (gure 14). Think about points such as which class would be more polar or non-polar, would the shapes and sizes of the molecules have any inuence? Note your comments in the grid below.
Triacylglycerols Similarities Phospholipids Steroids
Triacylglycerols Differences
Phospholipids
Steroids
The basic similarity is all three structures contain high proportions of carbon and hydrogen. This hydrocarbon dominates the triacylglycerol, with three long chain fatty acids. These are therefore the most non-polar and unreactive of the three, as anyone who has tried to wash fats and oils off dinner plates can say. The phospholipids have more mixed properties, enabling them to interact with both oils and water (we shall return to this point under Uses). In the steroids, the hydrocarbons are looped into a series of interlocking rings, giving a more compact molecule. This can pass though cell membranes.
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2.6 Uses of lipids Energy supply and storage Fats and oils are a rich supply of energy. They have caloric values of around 38 kJ/g, which is more than twice that of carbohydrates. Think of the difference between a plate of plain spaghetti (largely starch) and one with a rich sauce. Lipids are insoluble in water and therefore do not cause any problems with osmosis. They t the criteria we discussed above for polysaccharides as good storage molecules. Humans store 3-4 months supply of energy in the form of lipid, we tend to say fat. It is the main energy supply for the heart and for skeletal muscle for steady, sustained exercise. In contrast, our stores of glycogen would be exhausted by a marathon, a few days fast or a weeks slimming diet. If people are given free access to food and little need for physical activity, many store more lipid than is fashionable or the mortality statistics suggest is good for them. Other uses of stored lipids Stored lipids (fat) have other uses than energy supplies. Subcutaneous fat gives mechanical protection. Think of the experience of a blow somewhere where the layer of fat under the skin is thin, such as the shin. It also gives thermal insulation. Layers of fat contribute to body shape, particularly in women. Cell membranes Figure 13 described and illustrated the structure of phospholipids. Activity 12 extended this to consider their properties. They make good emulsifying agents. A double layer allows the hydrophilic parts of the molecules, such as the phosphate groups, to be attracted to water. In practice, this means the inside of the cell and the watery uids bathing it. The hydrophobic parts of the molecule, mainly the long hydrocarbon chains of the fatty acids, twist into the centre of the membrane. This gives a bilayer, the basis of a cell membrane. Waterproong Everyday experience shows us that covering things with a layer of grease keeps water out. It can be equally useful for keeping water in. Most surfaces inside the body have a layer of mucus, which is largely lipid, for waterproong and lubrication. This is important to anyone interested in Cosmetic Science because many products are designed to counteract the natural tendency of the skin to dry out. A good coat of goosegrease, worn daily, would do much to reduce water loss and challenge the signs of ageing. However, this would be cosmetically unappealing. A more pleasant texture can be produced by mixing the grease with water. Molecules with opposite properties at each end, such as phospholipids, can hold these together to form emulsions. These are explained elsewhere in the course. Phospholipids are also the main emulsifying agents in body uids such as the blood. Essential fatty acids Animals such as ourselves cannot produce the polyunsaturated fatty acids. They are therefore essential in our diet. Many of their body functions are nutritional and beyond the scope of this Unit. They are the basis for the production of important regulatory substances, such as the prostaglandins. A problem with polyunsaturated fatty acids, particularly when they are found in cell membranes, is that they are particularly susceptible to oxidising reactions. These trigger chain reactions, leading to rancidity. Both the food industry and the cosmetics industry control this with the use of preservatives known as antioxidants or free radical scavengers. Some of these are synthetic. Some are the substances naturally used by the body for the same purpose, such as the vitamins A, C and E. Hormones Some lipids, in the form of steroids act as hormones in the body. Examples are oestrogens (female hormones), androgens (male hormones) and cortisones. Steroids, being compact lipid molecules (Activity 12) can pass through cell membranes and have profound effects on their activities. Some other hormones are molecules which are water soluble and must work by attaching to the membrane and inuencing what can pass through it. Insulin is a good example, inuencing the way in which cells can take up sugars. An effect of steroid hormones of particular importance to the cosmetics industry is that they affect uid balance. For example, oestrogens (female hormones) tend to cause water retention. This helps to maintain womens skin. This effect reduces with age. The cosmetics industry is sensibly aware of this.
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2.7 Summary The lipids are a mixed group of bio chemical substances whose main similarity is that they will not mix with water. This is due to the presence of a high proportion of hydrocarbon in their molecules. Major types of lipid include triacylglycerols (fats and oils), waxes, phospholipids and steroids. Important biological functions of lipids include energy supply and storage, formation of cell membranes, waterproong, as nutritionally essential bio chemical metabolites and as the molecular basis for some important hormones.
CHECKLIST At the end of this section, you should be able to: describe the nature of lipids as biological molecules containing a high proportion of hydrocarbon, making them immiscible with water. describe and illustrate the structure of fats and oils as combinations between glycerol and three molecules of fatty acids. explain how the insertion of double bonds into molecules of fatty acids alters their shapes and therefore their physical and biological properties. compare and contrast the structures of a range of other lipid material, with reference to waxes, phospholipids and steroids. explain the biological roles of lipids with reference to energy supply and storage, cell membranes, waterproong and hormonal activity.
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APPENDIX 1 Answers to Activity 1 The eggs have been used as experimental models to show how osmotic gain or loss of water could affect normal body cells. You should have noted that eggs in the sugar solution shrank but that those in water swelled up and the membrane might even have burst? Your explanations should be: In the saturated sugar solution, glucose molecules were too large to pass through the cell membrane. Because there was less space for water in the sugar solution, the extracellular concentration of water was lower than the intracellular concentration. Water can move fairly easily through the egg cell membrane, therefore it diffused outwards down its concentration gradient and the cell decreased in volume. The reverse situation of course applied to the egg placed in water here, the intracellular uids of the egg white and yolk contain proteins and other solutes which are too large to easily pass through the membrane. They, however, decrease the space available for water molecules, i.e. the relative concentration of water is lower than in the extracellular solution, therefore water enters and the egg cells swell. At the end of the experiment, you should nd that the egg soaked in the sugar solution has gone quite rm. This is a result of the excessive loss of water and the protein has become coagulated or denatured. IMPORTANT NOTE The experiment illustrates the importance of maintaining a relatively constant balance of solutes (such as glucose, but also of ions) between intracellular and extracellular uids (especially the blood). The importance of homeostatic control systems will be reinforced during later discussions of the physiology of blood, the kidney, etc. If you dont understand these answers, revise the section in the text explaining osmosis.
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Notes
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3 PROTEINS 3.1 Introduction Proteins are the third class of biological molecules covered in this unit, and are the molecules within biological systems that do the work required for life. In order to relate the effects that cosmetics products may have on living systems, we need to understand something of the basic principles that govern protein structure and function. The goal of this section, therefore, is to introduce the principles of protein structure and outline the essential variety of functions that proteins perform within living systems. First, I will introduce amino acids, which are the basic building blocks of proteins, and then go on to look at how these building blocks are incorporated into proteins, and ultimately how they dene the threedimensional structure and function. Some of the principles that you learned in the Basic Chemistry unit will come in useful as you learn about proteins. 3.2 Amino acids General structure Amino acids are the basic building blocks of proteins and all have the same basic structure. They contain an amino group, a carboxylic acid group and a side chain group bonded to one carbon atom called the -carbon.
R CH amino group H2N
side chain group COOH carboxylic acid group
-carbon
Figure 15. General structure of amino acids
Humans have 20 amino acids which are genetically encoded. Of course, these follow the general structure outlined above, and differ from each other in the nature of the side chain group. We will look more closely at the different side chains later in this section. Amino acids are chiral. All of them (with the exception of glycine) have an asymmetric -carbon atom. This means it is bonded to four different groups, i.e., the amino group, the carboxyl group, the side chain and hydrogen. The -carbon atom is the chiral centre. The four groups can be arranged in two different ways around the chiral centre: one arrangement is the mirror image of the other. The two forms cannot be superimposed on one another and so are isomeric. The technical term for these two isomeric forms is enantiomers or optical isomers. ACTIVITY 7 Optical isomers of amino acids (Allow 5 minutes) A general structure of an amino acid is shown in gure 16. Draw its mirror image in the space provided.
H2N
Figure 16. Structure of an amino acid
COOH
mirror image
If you have any difculty, you could use the molecular model kit to represent one amino acid and then try to assemble its mirror image. The answer is in Appendix 1 at the end of this section.
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The side chain for glycine is hydrogen. You should note that as glycine has two hydrogens attached to the -carbon, it isnt chiral it is achiral. This is because with two of the attached groups being the same, you cant produce mirror images that are non-superimposable, the two forms are called the L and D isomers. The 20 genetically coded amino acids that occur in proteins are all the same single enantiomer and are given the designation L. The example in gure 17 shows Lalanine and D-alanine.
CH 3
CH 3 H
H2N
COOH
H2N
COOH
L-alanine
Figure 17. Structure of L-alanine and D-alanine
D-alanine
Single amino acids in aqueous solution are ionised and can act as acids or bases. At neutral pH, 7.0, the pH found within cells and maintained within the body, amino acids exist as a doubly charged zwitterion as shown in gure 18.
R pH 7.0 aqueous solution + H 3N
net charge =0
R COO
H 2N
COOH
net charge =0
Figure 18. Ionisation of amino acids
This is electrically neutral, net charge 0. Note, however that the net charge on specic amino acids also depends on the nature of the side chain. If the side chain is acidic or basic, that too will be ionised at neutral biological pH. Therefore, amino acids with acidic or basic groups will have a net ionic charge. Forces between ionic charges (electrostatic) are very strong, so interaction between charged side chain groups when incorporated into proteins is important in the maintenance of protein structure (as we shall see), and in providing the driving force for catalytic activity in some enzyme sites. Essential amino acids We have said that 20 amino acids are genetically encoded. This means that the machines within cells that synthesise proteins have the capacity to uniquely recognise each of these amino acids and incorporate them specically into their correct place within a given protein. Only nine of these amino acids are required in the human diet, i.e., our bodies cant make them. They are said to be essential. The essential amino acids are listed in table 2 later on, and include histidine, leucine and lysine. The remaining 11 amino acids required in proteins can be derived from the essential amino acids or synthesised from scratch using ingredients from the metabolic pathways within the cell. Side chain classication Amino acids can be broadly classied in terms of the properties of the side chain group. In particular, the polarity of the side chain greatly affects the properties of the amino acid. The polarity is the ability to interact with water at biological pH (7.0) and the side chains vary greatly from non-polar, hydrophobic (water hating) to highly polar, hydrophilic (water loving) groups. Polar amino acid side chain groups can further be divided into those which are ionically charged at biological pH and those which are electrically neutral. Other factors that vary between amino acids and side chains are size and shape. We will see later how these factors; polarity, size and shape of the side chain groups of amino acids are fundamental to determining the structure and function of proteins which are made up of amino acid building blocks.
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DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5 Table 2. Amino acid classication groups
General structure of amino acids Basic (polar, positively charged*) Non-polar

H2N
R COOH
Structure of R
CH2 CH2 CH2
Name
Abbreviation
Lys
Structure of R
H3C CH CH2
Name
CH3
Abbreviation
Ile
NH2 Lysine (E)
CH2
NH H3C CH3 CH CH2
Isoleucine (E) Leucine (E)
Leu
Arginine
CH2 CH2 CH2 N N H NH2
Arg
H3C CH
CH3
Valine (E)
Val
Histidine (E)
NH CH2
His Phenylalanine (E) Phe
Acidic (polar, negatively charged*)

COOH CH2 COOH CH2 CH2
CH2
Aspartic acid (Aspartate) Glutamic acid (Glutamate)
Asp
CH3
Alanine
Ala
Glu Tryptophan (E)

NH
Trp
Neutral (polar, non-charged*)

OH CH2
CH2 OH
Serine
Ser Tyrosine Tyr

CH2
H3C CH
OH
Threonine (E)
Thr
SH CH2
Cysteine
Cys
CH2 CH2
CH3
Methionine
Met
CONH2 CH2 CONH2 CH2 CH2
Special amino acids Asparagine Asn

HN
Proline
COOH
Pro
Glutamine * = at pH 7.0 (E) = essential amino acids
Gln
CH2 H2N COOH
Glycine
Gly
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Mention must be made of the two special cases, proline and glycine. Proline is the only genetically coded amino acid where the side chain is cyclised into the amino acid nitrogen to form a ring. The amino group is now a secondary amine as opposed to a primary amine in all the other amino acids. The cyclic structure of proline means it is somewhat less exible than the primary amines when it is incorporated into peptides and proteins. This has important consequences for the structure of proteins as we shall see later. The side chain group of glycine is hydrogen, which makes it the smallest amino acid possible. Because of this it is somewhat more oppy than other amino acids, and like proline, this can also affect protein structure in an important way. Another amino acid worth special mention is cysteine, which can exist in proteins in two forms. The amino acid cysteine contains a thiol group (sulphur containing) which can be oxidised in combination with another cysteine molecule to form a disulphide bridge as shown in gure 19. SH HS O2
+
cysteine cysteine cystine
Figure 19. Formation of a disulphide bridge by oxidation
The resulting bridged amino acid is something called cystine. These disulphide bridges between cysteines are a very important feature in protein structure. Despite the huge variation in properties that the genetically coded amino acids can confer on proteins, there are several examples of unusual or modied amino acids that occur in nature. Hydroxyproline is a major constituent of the connective tissue protein collagen, and is formed by the oxidation of proline. The modied amino acids are usually derived in the same way from genetically encoded amino acids. Amino acids with a D conguration are found in some lower organisms, particularly bacteria, as constituents of cell walls or antibiotics. The Dconguration acts as a defence mechanism because it is resistant to metabolic and hydrolytic enzymes.
ACTIVITY 8 Inuence of amino acid side chains (Allow 10 minutes) The amino acid side chains play a fundamental role in determining the properties of an individual amino acid. It is therefore worthwhile recognising the side chains and their likely inuence.
CH3 CH2 CH2 CH3
CH2 CH2
CH2 NH2
COOH CH2
CH2
OH
4
Figure 20. Amino acid side chain groups
CH2
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Shown in gure 20 are 5 amino acid side chain groups. Assign these side chain groups to the appropriate classication groups listed below. 1. 2. 3. 4. 5. Side chain classication groups A. Non-polar B. Neutral, polar C. Basic D. Acidic E. Special
The answers are provided in Appendix 2. 3.3 Peptides Molecules of amino acids can be linked via a condensation reaction to form a polymeric chain of amino acids called a peptide. When two amino acids are condensed together, this forms what is known as a dipeptide (see gure 21). We saw the same type of reaction in the formation of polysaccharides from monosaccharides.
R1 OH H2N C O H H N R2 H2O Figure 21. Condensation reaction to form a dipeptide COOH H2N O R2 R1 H N COOH
The condensation takes place in this instance between the amino group of one amino acid molecule, and the carboxylic acid group of another amino acid molecule. The reaction involves the loss of water (hence condensation) and forms an amide bond (see gure 22). In peptides this amide bond linkage is known specically as a peptide bond. R1 N O
Figure 22. Structure of a peptide bond
H N R2
The peptide bond is planar, i.e., the oxygen, carbon and nitrogen atoms lie in one plane. Once a dipeptide has been formed, another amino acid molecule can be condensed with a dipeptide to form a tripeptide.
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ACTIVITY 9 Formation of a tripeptide (Allow 10 minutes) Earlier you saw how carbohydrate molecules could combine to form a bigger molecule. Amino acids can also form larger molecules via the process of condensation. To illustrate, try to draw the resulting tripeptide if the dipeptide and amino acid shown below join together via a condensation reaction.
amino acid dipeptide
O OH H2N C O
phenylalanine
H2N CH3
OH N H C O
alanineleucine
The answer is provided in Appendix 2.
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Of course you could add a fourth amino acid via condensation to form a tetrapeptide, and a fth to form a penta-peptide, and so on. This process can continue to form chains of peptides of varying length and order of amino acids. These are known collectively as polypeptides. Polypeptides of sufcient length may become a protein. However, a protein is not just a long polypeptide! Key differences between a polypeptide and a protein A polypeptide is a single chain of amino acids with varying length as we have seen. It need not have a dened three-dimensional structure. It also need not have a biological function. A protein may however, consist of one or more polypeptide chains. Proteins are very large polypeptides with greater than 50 amino acid residues. They have a dened three-dimensional structure, and all proteins have a distinct biological function. When condensed into a peptide chain, the portion of the amino acid that remains after condensation, i.e., the side chain, the nitrogen and the carbonyl group is known as the amino acid residue.
polypeptide backbone
OH O
N-terminus
H2N O
N-terminal residue (leucine)
H N
N H O
serine residue
H N
COOH
C-terminus
alanine residue
C-terminal residue (phenylalanine)
Figure 23. Key features of a polypeptide chain
This term is used quite often to describe the side chain groups within a polypeptide chain. A polypeptide chain contains two amino acid residues which are only partially condensed one at each end. These are the terminal residues. The residue at the end with the free amino group is called the N-terminal residue or N-terminus, and the residue at the opposite end that has a free carboxylic acid group is called the C-terminal residue or C-terminus. The chain of atoms that comprises the nitrogen, -carbon, and carbonyl carbon is referred to as the peptide backbone. In peptide shorthand, using three-letter codes, the peptide chain is always written starting from the N-terminus. Thus the tetra-peptide example in gure 23 would be written Leu-Ala-Ser-Phe. When condensed into a peptide bond, the nitrogen of the amino group is no longer basic, and the carboxyl group is no longer acidic. This means that the properties if a peptide are very much dependent on the collective properties of the side chain groups of the amino acid residues.
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3.4 Proteins As we mentioned earlier, proteins are the molecules within biological systems that do the work required for life. They may need instructions on what to do from genetic material (DNA, see later). They may need energy to carry out their functions (from metabolism), but only proteins are capable of the enormous diversity of structure and function required to maintain life. Because of this, proteins are difcult to classify. The most useful classication is by their function and even this is very broad. Within each functional group, there is a vast variation in protein properties as is outlined below: Table 3. Protein properties
Protein type Structural protein Enzymes Contractible proteins Transport proteins Regulatory proteins Protective proteins Storage proteins Genetic protein Example and function Keratin, armour-like protein found in hair, skin and nails These are biological catalysts and carry out the chemical reactions on which life depends. They will be covered separately later. Myosin and actin, the proteins responsible for muscle contraction Haemoglobin, responsible for the transport of oxygen in blood Hormones such as insulin which regulates glucose levels in the body Antibodies which recognise foreign substances in the body and help to eliminate them Ovalbumin, the major protein of egg white Histones which help organise the structure of DNA within the cell nucleus
A complementary way of classifying proteins is on the basis of their shape. Globular proteins have their polypeptide chains folded into compact shapes and are nearly always soluble in aqueous solution. Examples of globular proteins are enzymes and antibodies. Fibrous proteins have their polypeptide chains extended on one axis rather than folded, and are usually insoluble in water. They tend to serve a structural role. Typical examples are keratin in hair and collagen in connective tissue. You may think that a long polypeptide chain consisting of single bonds can arrange its atoms in an innite number of ways in three-dimensional space. The three-dimensional arrangement of atoms within a molecule is called the conformation. Proteins do have dened structures, however, which determines their biological activity. Furthermore, proteins can be isolated without losing their biological activity thus implying that the conformation of the polypeptide chain within proteins is quite stable. We shall see how proteins achieve this stable conformation by considering the levels of protein structure. 3.5 Protein structure We have distinguished proteins from simple polypeptides and said they have a dened structure. The structure of proteins is complex and has several different levels. Each of these levels of structure is built upon the previous one in a hierarchical manner, increasing in complexity and scale at each stage, starting with the order of amino acids in the chain, through to the overall three-dimensional structure of the molecule.
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Primary structure The rst level is the primary structure. This is simply the amino acid sequence of the protein. It can also include the location of the cysteine residues that form disulphide bridges. To use the enzyme ribonuclease as an example, the sequence and location of the disulphide bridges is shown in this diagram.
10 Glu Arg Gln His Met Lys Phe
Ala Thr 1 Lys +H3N 30 Lys Ser Arg Asn Leu Thr Lys Asp Arg Ser Lys Tyr Pro Asn Cys Ala Cys Lys Pro Val Asn Tyr 124 Val Ser Gly 90 Met Met Gln Asn Glu
Ala
Ala
Asp Ser
20 Ser Asn Tyr Cys Cys Arg Asp Ala Ser Thr Ser Ser Ala
Ser
80 Thr Ser Tyr Ser Gln Tyr Ser Met Ile Cys Thr Asn 70 Thr Gln Lys Asn Gly Cys Ala Val Asn Glu Lys Gln Ser Cys Val Ala Gln Val Asp 60
Thr Glu 120
Ser Ala Asp Phe His Val Pro Val Tyr Pro Asn Gly C 0Val Lys 100 His Lys Thr Thr Gln Ala Asn Ile Ile 0 110 Ala
Cys
Thr Phe Val His Glu Ser Leu 50
Ala
Figure 24. Primary structure of the enzyme ribonuclease showing location of disulphide bridges
As can be seen, this gives more information about the three-dimensional structure of the protein and shows that just because amino acid residues can be well separated in their sequence along a peptide chain they may well be close in space. Serine-59 and Glutamic acid-111 in ribonuclease are separated by 50 residues, but because of the presence of the disulphide bridge adjacent to them they are held much closer in space and their actual separation is more like two residues. Secondary structure The next level of protein structure is secondary structure. Here we will consider short sections of the overall chain which have localised structure. It is a bit like knots in a piece of string. Primary structure is the string itself. Secondary structure is the type of knots in the string, and we will go on to consider how the ball of string coils round on itself later. The favoured conformations of the polypeptide chain lead to two main types of secondary structural elements although there are others. These are the -helix and -sheet. The -helix is shown in gure 25.
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H C C N
N O
H N C
C O
C O
H N C
C O C H N H N O
C C O
H N
H N
C
H N
C O
C
H
O
C
H C
C
C
N
H N
O
C
N O
H C O N C
C O
Figure 25. Representation of the -helix of a polypeptide chain
This shows the helical arrangement of -carbons and the stabilisation of the helix by hydrogen bonds. The carbonyl group of each amino acid is bonded to the N-H of the peptide bond of the residue four ahead of it in the linear sequence, and all of the carbonyl and N-H groups in the peptide chain or backbone participate in hydrogen bonds. In some structural proteins, lengths of -helix can entwine around each other in a cable-like structure called a -helical coiled coil, for example, keratin in hair. Shorter lengths of -helix can occur as structural elements of globular proteins such as enzymes. We shall see this later. Breaking and re-forming disulphide bridges in hair keratin is the basis of perming where hair is formed into appropriate curls. A solution of a reducing agent is applied with heat. This serves to break the disulphide bridges into the thiol groups of cysteine. The damp heat disrupts the hydrogen bonds on the -helices and causes them to uncoil and stretch. After the reducing solution is rinsed off, an oxidising agent is applied. This reforms the disulphide bridges between cysteines but not in their original positions. The hair bres reform the -helices but the new disulphide bridges constrain the bre and produce the desired curl. The second main secondary structural element in proteins is the -pleated sheet or -sheet more commonly. As in the -helix, the peptide bond participates fully in hydrogen bonding. However, the bonding is between different polypeptide chains, or inter-chain, rather than between the same polypeptide chain or intra-chain.
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The major difference between the -helix and -sheet is the conformation of the peptide chain backbone which is in a fully extended conformation rather than a helical structure. The polypeptide chains in a -sheet can run in the same direction as each other a parallel -sheet, or in an opposite direction antiparallel -sheet (see gure 26).
N C C N H O C O C
N C O H N
C N H O C
C C H N
C C C
N C C N H O C O C
N C O H N
C N H O C
Figure 26. Parallel and anti-parallel -sheets
This type of structure is found in brin, the protein of silk and spiders webs. It is more exible than keratin but cannot be stretched. As the side chain groups stick out of the plane of the -sheet structure (not shown in the diagram), in general only small amino acid residues are compatible with this type of structure, e.g., alanine or glycine. Shorter elements of -sheet secondary structure can be found in globular proteins along with -helices. Collagen, the tough, brous connective tissue has an extremely high proportion of proline, glycine and hydroxyproline in its structure. As such, -helices and -sheets are not available as secondary structural elements. Another structural type, the collagen helix is found instead and is unique to collagen. The elastic connective tissue protein elastin has yet another type of helical secondary structure that is neither an -helix or a collagen helix. We shall see later how the collagen helices are arranged when we discuss higher levels of protein structure. We shall also compare this with the structure of elastin. 3.6 Tertiary structure This is the third level of protein structure and is concerned with the gross three-dimensional structure of a polypeptide chain and three-dimensional interrelationship between elements of secondary structure within a polypeptide chain. The shape a particular protein adopts is dependent upon a large number of relatively weak interactions between amino acid side chain groups. Disruption of these weak forces is relatively easy and can be done by heat, changes in pH or by heavy metal ions. The disruption of the three-dimensional structure of protein is denaturation. An everyday example of this can be seen whenever we cook an egg.
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Egg albumin, the protein of an egg white, changes on heating from a thick clear solution of globular protein to a congealed mass that cannot be returned to its clear soluble state. It is said to be irreversibly denatured. Hair perming as we saw earlier is a rather more controlled example of protein denaturation. When a protein is denatured it loses its biological activity. The dened three-dimensional structure is lost and the protein assumes a random coil conformation. Under controlled conditions some proteins can be denatured reversibly and returned to its original or native state (see gure 27). This is an important technique in biochemistry for purication and isolation of proteins from mixes of other proteins. As an example we shall return to the enzyme ribonuclease.
1. Reduction of disulphate bridges 2. Denaturation using 8M urea solution 72 65 84 95 84 95 40 110 110 58 1. Removal of urea by dialysis 2. Air oxidation of thiol groups Denatured Random Coil no enzyme activity SH SH 26 SH SH SH SH SH 58 SH 40
72 26 65
Ribonuclease Native State cysteine disulphide bridges
Figure 27. Denaturing of ribonuclease and its reversal
Breaking the disulphide bridges in a controlled manner removes the constraints on the polypeptide chains. Concentrated solutions of urea act as a denaturing agent by disrupting the non-covalent weak forces that hold the peptide chain in its native conformation. The random coil ribonuclease is now a simple polypeptide; it has no enzymatic activity. The activity can be recovered by removal of the denaturing agent by a process called dialysis. This is achieved by placing the denatured protein solution into a semi-permeable membrane that allows the small urea molecules to diffuse out but leaves the large peptide chain inside. Reoxidation of the thiol groups to disulphide bridges reform the native structure and allows complete recovery of the full enzymatic activity. This shows that the folded conformation adopted by ribonuclease in its native state is the most energetically favoured out of many. This is generally true in all single polypeptide chain proteins. It also shows that the information required to specify the threedimensional structure and its function is contained in its amino acid sequence. Thus when a protein is synthesised from instructions contained in the genetic code in DNA of a cell, the link between the genetic code for that protein and its function is contained in its amino acid sequence. A single change to one amino acid in the sequence of a protein may be enough to disrupt the structure critically and cause catastrophic losses of protein function. This is how the damaging effects of genetic mutations manifest themselves. 3.7 Forces involved in maintaining tertiary structure Covalent bonds These are the strongest of the forces involved. Covalent bonds between side chain groups of residues occur less frequently than other, weaker forces. They are not responsible for forming tertiary structure, but are formed after the polypeptide chain has folded and help to stabilise this initially formed three-dimensional structure. The most important covalent bonding is the disulphide bridge. (Other forms of covalent link are found in structural proteins such as collagen and elastin. These will be discussed later).
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Ionic This type of link occurs between side chain groups that have permanent electric charges at biological pH, e.g., arsinine, lysine, glutamine and aspartic acid. (See gure 28) These are sometimes called salt bridges.
NH2 Arg N H NH2
O Glu O
polypeptide chain
polypeptide chain
Lys
NH3
OOC
Asp
Figure 28. Ionic linkages
Hydrogen bonding We have seen how hydrogen bonding between peptide bonds in the polypeptide backbone helps to maintain secondary structure. Hydrogen bonding between side chain groups of amino acid residues is very important for the maintenance of tertiary structure. All amino acids with polar side chain groups are capable of hydrogen bonding with each other. Additionally, the hydrophobic amino acid residues tyrosine and tryptophan have groups that are capable of forming hydrogen bonds. Hydrogen bonds are also formed between polar groups and water. In soluble proteins it is common to nd polar and charged groups facing the exterior of the protein. Hydrophobic interactions Hydrophobic side chain groups (e.g., those of phenylalanine, leucine, valine, etc.) tend to be more stable when closely packed together at the centre of a globular protein than when they are exposed to water on the exterior. This is a small-scale molecular analogy to the immiscibility of oil and water. Protein chains will therefore tend to fold spontaneously so that the hydrophobic residues are buried in the centre and polar and charged groups face the outside. Van der Waals forces Atoms of side chain groups in tightly packed polypeptide chains are nearly all at a minimum distance apart so they maximise the Van der Waals force between them. The nal tertiary structure adopted by a protein develops by folding of the polypeptide chain. Elements of secondary structure; -helix, -sheets etc., develop spontaneously in a chain and organise themselves relative to each other driven by the weak forces of hydrophobic interactions, Van der Waals forces and nally hydrogen bonding. The nal three-dimensional conformation is locked by covalent bonds such as disulphide bridges.
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To illustrate the tertiary structure of a protein and its composition of secondary structural elements, examine the diagram of the enzyme carbonic anhydrase (gure 29). Note the -helices represented by helical ribbons and -sheets represented by at arrows. Note that the threedimensional structure precisely positions the three key histidine residues at the active site of the enzyme.
Histidine residue at active site
AcNH
-helix
-pleated sheet CO2

Figure 29. Structure of carbonic anhydrase
3.8 Quaternary structure If a protein consists of more than one polypeptide chain, quaternary structure is the term used to describe how the chains are packed together. Each polypeptide chain is called a subunit and each of the subunits has its own secondary and tertiary levels of organisation. Several forces stabilising the quaternary structure are the same as those found in tertiary structure i.e., disulphide bridges and weak intermolecular attractive forces. Multi subunit proteins are large. Examples include haemoglobin, the oxygen carrying protein of the blood (4 subunits; molecular weight 64,500); glutamate dehydrogenase a metabolic enzyme (6 subunits; molecular weight 320,000); and pyruvate dehydrogenase complex, a metabolic enzyme system (72 subunits; molecular weight 4,600,000). Collagen and elastin We shall now look at the structure of collagen and elastin. We saw earlier how collagen and elastin have their own specic secondary structural elements as a result of their unique amino acid compositions. In collagen, the helices are intertwined into a triple helix and these in turn are bound together into long inextensible rod-like bres by a unique system of covalent cross-links. (See gure 30.) These covalent bonds are stronger than disulphide bridges and help to provide the high tensile strength of collagen, e.g., in cartilage.
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Short section of bre a collagen bre
Collagen molecule
Collagen triple helix
Figure 30. Diagrammatic representation of the structure of collagen
Elastin has a much looser organisation. The short lengths of polypeptide helix are folded and twisted and have a system of covalent cross links between polypeptide chains similar to collagen, but it allows elastic deformation of the structure without breaking any of the bonds. (See gure 31.)
Crosslink
Stretch Single elastin molecule Elastic fibre

Figure 31. Structure of elastin illustrating elastic properties of the protein
Relax
Elastic fibre
This is important in tissues such as the lungs and arteries where strength but elasticity are required to cope with expansion and contraction.
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ACTIVITY 10 Levels of protein structure (Allow 10 minutes) A major determining feature of protein structure relates to the bonds and forces that maintain their three-dimensional structure. Here is an opportunity to conrm your understanding of this relationship. Based on what you have learnt, complete the following table:
Level of structure Primary Bonds and/or forces involved
Secondary
Tertiary
Quaternary
The answers are in Appendix 2 at the end of this section.
3.9 Summary The structure and function of a protein is determined by the properties of the sidechains of its sub-units, amino acids, which are linked together by peptide bonds which form via a condensation reaction. As more amino acids are joined to the chain, a polypeptide is formed. Proteins are made up of one or more polypeptide chains and have a dened three-dimensional structure. They also have a biological function. The structure of proteins is made up of four levels: primary the order of the amino acids in the polypeptide chain secondary the localised structure of polypeptide chains held in place by hydrogen bonds tertiary the three-dimensional relationship between elements of secondary structure held in place by various forces quaternary the overall three-dimensional structure formed by more than one polypeptide chain in a given protein.
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CHECKLIST At the end of this section you should be able to: draw the basic structure of an amino acid and name its constituent parts explain the principle of chirality as it applies to amino acids dene essential amino acids describe the different classications of amino acid side chains draw the reaction scheme for two amino acids (or amino acid residues) condensing to form a peptide bond, and identify the peptide bond dene a protein and the different functions it can perform describe primary, secondary, tertiary and quaternary structures of proteins, to include: understanding the principal forces involved in holding the various levels of structure in place understanding the structural elements present in each level of structure, e.g., -helices, and some of their properties compare the structure of collagen and elastin and relate this to their different functions as connective tissue.
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4 ENZYMES 4.1 Introduction As we have seen already, one of the most important functions of proteins is as enzymes: their structure and function is vital to our biological processes. The word enzyme comes from the Greek, en in and zyme yeast. This reects the fact that a lot of the knowledge we have today was obtained by studying yeasts and other micro organisms. Enzymes catalyse chemical reactions, typically speeding them up by 108-1010 fold, but it can be more! The power to catalyse reactions far exceeds all man-made catalysts. Man has been using the properties of enzymes for many centuries to produce such things as alcohol and cheese. In recent years, the uses of enzymes have become much more sophisticated with enzymes being used to produce chemicals for the pharmaceutical industry and as active ingredients in washing powders to attack protein stains. 4.2 Function of enzymes A major function of enzymes is to act as catalysts. However, a more complete denition would be: Proteins of biological origin which increase the rate of specic reactions without affecting the nal position of the reaction equilibrium. Enzymes catalyse reactions by reducing the amount of energy needed to convert compound A through a transition state T of higher energy, to compound B (see gure 32 below).
T
Activation energy of uncatalysed reaction Activation energy of enzyme catalysed reaction
Reaction A B
Energy
B Path of Reaction
Figure 32. Reaction pathway of an enzyme catalysed reaction
The enzyme reduces the amount of energy, activation energy needed to convert A to B, so making it easier to convert A to B. It also makes it easier for B to be converted to A by the same degree, so the position of the reaction equilibrium remains the same. Enzymes do this selectively, in fact, enzymes are very specic such that any one enzyme only catalyses one reaction or set of closely related reactions.
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4.3 Enzyme classication Classication of enzymes became necessary because of their vast number and variety, and also because of inconsistencies in the nomenclature that had developed over the years. A useful rule of thumb is that most enzymes have names that end in -ase, with the main exceptions being proteolytic enzymes such as trypsin ending in -in. Furthermore, the names of enzymes may to relate to the substrate involved. So a lactase enzyme has lactose as its substrate. The lactase enzyme hydrolyses the lactose, breaking it into glucose and galactose. So what do you think a fumarase enzyme would do even if you had never heard or seen it? You would be right to think that the substrate was fumarate, but this reaction is not a hydrolysis but a hydration reaction, the addition of water to produce maleate. It is unfortunate that the type of name sometimes tells you nothing about the type of reaction it catalyses, merely the substrate. However, some enzyme names tell you what type of reaction is involved. A transcarboxylase enzyme catalyses a transcarboxylation reaction. But in this case the name tells you nothing about the substrate. It is because of this confusion that a systematic way of naming enzymes was developed. We wont go into great detail here, but just look at some of the principles of this system. The Enzyme Commission divided enzymes into six classes with the basis for the division being the total reaction the enzyme catalyses. The number (1-6) of the class is then the rst digit of a fourdigit code assigned to an individual enzyme.
ACTIVITY 11 Classication of enzymes (Allow about 5 minutes) In order to help you get a feel for the classication of enzymes, complete the following exercise. Listed below are the classications of enzymes, and the denition of the classes. Match the letter of the denition with the appropriate number code for the classication that matches it. This will help you to get a perspective of the vast number of different reactions that enzymes catalyse, and how they are grouped together.
A B C D E F Hydrolysis reaction Isomerisation reaction Oxidation and reduction reactions The synthetic joining of 2 molecules coupled with the breakdown of a pyrophosphate bond in a nucleoside pyrophosphate Removal of a group from a substrate (not by hydrolysis) Transfer of an atom or group between two molecules (as long as it isnt covered by another of the classes).
1. Oxidoreductases; 2. Transferases; 3. Hydrolases; 4. Lyases; 5. Isomerases; 6. Ligases You can nd the answers to this activity in Appendix 3.
The rest of the four-digit classication code is made up as follows. The second and third digits in the code describe in more detail the type of reaction catalysed. The fourth digit describes the substrate. The structure of an enzyme will determine how it acts on its substrate.
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4.4 Enzyme structure As we have seen previously, enzymes consist of protein chains, either just one chain or a combination of chains. Although these chains can be of great length, only a few specic amino acid residues will actually be involved in the function of a specic enzyme. The other amino acid residues in the chain create and hold together the three-dimensional structure of the enzyme, which is critical to its role as a catalyst. The substrate is effectively the starting material of the reaction, which will eventually be converted to the product. The enzyme and the substrate form a complex which is the key to the catalytic activity of the enzyme. Once the reaction has taken place, the product is released from the complex and the enzyme is ready to receive a new substrate molecule (see gure 33).
Enzyme
Substrate
Enzyme substrate complex
Enzyme
Product
Figure 33. The process of enzyme catalysis
4.5 The enzyme-substrate complex The site on the enzyme where the substrate is bound is called the active site. This consists of a cavity formed by a specic arrangement of amino acids on the protein surface. These amino acids do not have to be adjacent to each other in the protein chain; they may be from separate parts of the chain, but the three-dimensional structure of folds etc. brings the required amino acids close together as we have seen earlier. These amino acid residues are only a fraction of those in the chain, and these are held together by the rest of the chain. The enzyme hen-egg lysosyme cleaves certain bonds between residues in the polysaccharide components of bacterial cell walls, so maintaining sterility within the egg. The schematic diagram of lysosyme shown here has the 19 amino acid residues that are part of the active site marked. You can see that they form widely separated clusters along the chain. When the protein chain is folded into its three-dimensional structure as shown in gure 34, there is a cleft (arrowed) containing the active site, where the polysaccharide chain ts and is distorted as it binds to the enzyme. The enzyme also changes shape when it is bound. A variety of interactions stabilise the binding.
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+H3N COO -
Figure 34. The three-dimensional structure of hen egg lysosyme
ACTIVITY 12 Enzyme structure and mechanism (Allow about 5 minutes) You already know something about the forces involved in determining protein structure. Apply what you have learnt to the following to help increase your understanding of enzyme function. A) Bearing in mind the various properties of amino acid residues you have learnt about, list three forces or types of bonds you think may be brought into play at the active site of an enzyme as it binds a substrate. For example, you may have suspected that hydrogen bonding would play a part, and you would be right. 1.
2.
3.
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B) There are two ways that enzymes and substrates t together: one is called induced t, the other is called lock and key. Figure 35 shows schematic diagrams of both mechanisms. Assign the correct mechanism to the diagrams, and explain what you think happens in each case as the complex is formed.
Substrate
Substrate
Enzyme
Enzyme
Enzymesubstrate complex
Figure 35. Formation of the enzyme/substrate complex
You can nd the answers to this activity in Appendix 3. When the substrate molecule is bound in the active site of the enzyme, the reactivity of the substrate is increased. The substrate may undergo a small change in shape so that it is put under strain to make it more reactive. Another method is for the enzyme to form a covalent bond with the substrate. In the case of some of the proteases (e.g., chymotrypsin), a charge relay system activates a serine residue, which in turn binds with the substrate, hydrolysing a peptide bond. Sometimes in addition to the properties of the amino acids at the active site, enzymes need the presence of coenzymes or cofactors to help them catalyse reactions. Coenzymes are nonpolypeptide molecules which are bound to a greater or lesser extent to the enzyme protein. Coenzymes can be very complex molecules. Some are changed by the reaction they are involved in, others are not. Cofactors perform a similar function to coenzymes and are often metal ions. An example of a coenzyme is NADP (nicotinamide adenine dinucleotide phosphate) which acts as a coenzyme in oxidation/reduction reactions. Once the enzyme has catalysed the reaction, the product molecule is released and the enzyme will then bind the next substrate molecule and convert that into product and so on.
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4.6 Enzyme inhibition Enzyme activity can be affected in a variety of ways, by bodily control and chemical means. The areas we shall look at here are irreversible and reversible inhibitors. 1. Irreversible inhibitors Irreversible inhibitors become covalently bound to the enzyme preventing the enzyme substrate complex being formed. Once bound to the enzyme, the effects of the inhibitor are as their name suggests, irreversible. Examples of this type of inhibitor include metabolic poisons such as nerve gas, so you can see that the consequences of irreversible inhibition of an enzyme can have catastrophic effects for the organism in which it operates! Active-site-directed inhibitors are substrate analogues, i.e., their structure is very similar to the structure of the usual substrate of the enzyme. These inhibitors bind directly and irreversibly to the enzymes active site or very near to it. For example, BrCH2CHO is an active-site-directed inhibitor for an alcohol dehydrogenase where the usual substrate is CH3CHO. You can see the similarities between the two compounds. 2. Reversible inhibitors Reversible inhibitors bind non-covalently to the enzyme, establishing a rapid equilibrium between the inhibitor and the enzyme. There are two types competitive and non-competitive inhibitors. Competitive inhibitors bear a resemblance to the substrate of the given enzyme, and compete directly with the substrate at the active site. Their effect can be counteracted by swamping the inhibitor with a large excess of substrate to vastly increase the substrate concentration. In contrast, non-competitive inhibitors do not bind at the active site, so dont compete with the enzymes usual substrate. Therefore, increasing the substrate concentration has no effect on the inhibition; the inhibition will only be removed when the inhibitor is removed.
ACTIVITY 13 Effect of different inhibitors (Allow about 5 minutes) It is important to understand what effect the different types of enzyme inhibitors have on reaction rates. In order to learn more about his, complete the following exercise. First, an example. Figure 36 is a graph of what happens to the rate of a reaction when an excess of irreversible inhibitor is added to the reaction mixture at time X. Clearly, the reaction stops when the inhibitor is added, and in this case enzyme activity is stopped for good. (NB. Vmax is the maximum rate of reaction).
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Vmax
Velocity of Reaction
Time
Figure 36. Effect of an irreversible inhibitor on reaction rate
Now draw graphs on the spaces provided to represent the following:

1. Competitive inhibition (left graph) 2. Non-competitive inhibition.
At time x, a slight excess of inhibitor is added. At time x, a slight excess of inhibitor is added. At time y, a large excess of substrate is added. At time y, a large excess of substrate is added. What happens to the reaction velocity at both these time points? At time z, the inhibitor is removed. What happens to the reaction velocity at these time points?
Vmax
Vmax
x Time
x Time
Notes:
You can nd the answers to this activity in Appendix 3.
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4.7 Summary Enzymes are proteins, and play a fundamental role in catalysing biological reactions. They form complexes with the substrate molecule and convert it to the product before releasing it and beginning to transform another substrate molecule. The t between the enzymes active site and the substrate is critical in the catalysis of the reaction. Enzymes can be inhibited by a variety of mechanisms, including reversible and irreversible inhibition.
CHECKLIST At the end of this section, you should be able to: describe the function of enzymes in biological systems understand the principle of enzyme classication describe the basic mechanism by which enzymes catalyse reactions describe the basic structure of an enzyme including the active site and how this relates to enzyme function describe the function of coenzymes and cofactors describe the lock and key and induced t models of enzyme-substrate complexes dene the different types of enzyme inhibition and draw graphs to show their effect on reaction rate under differing conditions.
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5 METABOLIC PATHWAYS 5.1 Introduction In this section we are going to look at metabolic pathways, the means by which cells break down extremely complex chemical processes into manageable portions. For this, you will need the CD, diagram E (below), and the two overlays that go with diagram E. This section of the CD covers a description of what a metabolic pathway is, and covers a specic example in detail. The example used is respiration. When you are ready, turn on the CD and start to learn about metabolic pathways.
CARBOHYDRATE
CYTOPLASM
GLYCOLYSIS
Hexose diphosphate
2 x Triose phosphate
2 x Pyruvic acid CO2
MEMBRANE
MITOCHONDRION
Acetyl CoA
Oxaloacetate
Citrate
TCA CYCLE
Succinate Oxo-glutarate
CO2
CO2
Diagram E. Outline scheme for cell respiration
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DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5 Diagram F. The chemical products of respiration (to accompany the CD Metabolic pathways)
The products of respiration give, from one molecule of glucose C6H12O6, a total of: 6 molecules of carbon dioxide 4 molecules of adenosiine triphosphate (ATP)* 10 molecules of reduced hydrogen carrier, NADH2 2 molecules of reduced hydrogen carrier, FADH2 The hydrogen atoms shall later be reacted with oxygen to produce water, which is excreted.
* 2 (net) from glycolysis and 1 from each turn of the TCA cycle
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6 GENETICS AND CELL REPLICATION 6.1 Introduction In this unit, we are considering key biological molecules and their function. This section gives an introduction to the molecules involved in passing genetic information from one generation to the next on the molecular level, and how they are also involved in production of proteins in the body. You will learn about the structure of these vital molecules and how (like other molecules you have already studied) they are made up of simpler sub-units, as well at looking at how they are produced in the body and their function. We can all see evidence of inherited characteristics all around us, for example, in our families and when a seed produces a plant that is the same as the parent plant the seed came from. As early as the 19th century it was known that information was inherited via the chromosomes, which are seen in the cell nucleus as the cell begins to divide. But the discovery that deoxyribose nucleic acid (DNA) was the molecule that made up genes rst came in the 1940s. The structure of DNA was hypothesised by Watson and Crick in 1953, and has since been proven by x-ray analysis and bio chemical research. Research has demonstrated that from inside the cell, DNA directs what happens as an organism develops, and can also copy itself almost indenitely. 6.2 The structure of DNA You may well have heard of the famous double helix structure of DNA, two chains wrapped around each other in a spiral conformation. Before we go on and look at the overall structure of DNA and its function in more detail, it is necessary to understand something of its basic chemical structure. Despite DNAs vital role in nature, it is a molecule made up of fairly simple units linked together which go to make up a huge molecule with innite variety. DNA consists of four basic subunits which are all classied as nucleotides. A nucleotide must contain the following parts, and those in DNA are no exception, as shown in gure 37.
OO
-
R O CH 2 H O H
a nitrogenous (nitrogen containing) base
P O
phosphoric acid (or phosphate)
H OH H
a pentose sugar (5 carbon atoms)

Figure 37. Basic structure of a nucleotide
The four subunits or nucleotides of DNA differ in the bases they contain. The structure of the four bases are shown in gure 38.
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NH2 C
Cytosine (C)
O
H
Guanine (G) C C
N
C
C C
H
H
N
C
N
C H
N
R
H2N
N
R
O
H C
Thymine (T) C C
NH 2
C
Adenine (A)
N
C
CH 3
H H
N
C
C C
N
C H
N
R
N
R
Figure 38. Structure of the bases found in DNA
It isnt necessary to learn these structures off by heart, but you do need to know that C and T are pyrimidine bases and A and G are purine bases and can be represented as shown in gure 39.
cytosine and thymine R
Figure 39. Representations of DNA bases
R = rest of nucleotide molecule R
guanine and adenine
The nucleotides are joined together via a bridge formed by the phosphate group; this is called a phosphodiester linkage (see gure 40).
CH 2 H O H
Base
nucleotide 1
H
phosphodiester linkage
H O H CH 2 H O H
nucleotide 2
P O Base
H O
-
H O H
P O
Figure 40. Phosphodiester linkage
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The sugar (deoxyribose) and phosphate groups alternate to form the backbone of DNA while the four bases form the side groups. The structure of deoxyribose is shown in gure 41.
R C H2 H O H
H OH
Figure 41. Deoxyribose
H H
ACTIVITY 14 Basic structure of DNA (Allow about 5 minutes) Now you know all the parts of the DNA chain, see if you can draw a chain which includes all four bases joined together by the phosphate/sugar backbone. Remember, you dont need to draw out the full structures of the bases, just use the representations you saw earlier. To help, I have drawn out one of the nucleosides (adenine) now add the others!
O OP O O A CH 2 O
See Appendix 4 at the end of this section for my answer diagram.
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As I mentioned earlier, DNA has what is called a double helix structure. Two strands of DNA are wrapped round each other in a left-handed spiral as represented below (gure 42).
T A
C G
G C
C G
T A
T A
C G
G C
T A
A T
Bases
Sugar Phosphate Backbone
Figure 42. The DNA double helix
As you can see, all the bases (A, T, G, C) are inside the double helix, and the polar sugar/ phosphate backbone is on the outside of the structure. This makes sense because we have already seen that the backbone is polar due to the negative charge on the phosphate groups and the oxygen group in the sugar. Therefore, it is happy to sit in an aqueous medium, while the bases are hydrophobic (water-hating) and are kept inside the helix away from the water. Because of the structure of DNA, the bases have to be very close together in the double helix. In order for this to be possible, they form complementary base pairs. Although there are many ways in theory that the four bases could pair up, they only ever form the following pairs known as complementary base pairs. Adenine always pairs with thymine and cytosine always pairs with guanine. Each pair of bases is a combination of a purine (two rings) with a pyrimidine (one ring). This allows the two strands of DNA to always be the same distance apart. DNA and heredity Genetic information needs to be passed precisely from one cell to its offspring. DNA bases represent how our genetic code is spelt out via the bases A, T, G and C. The sequences of bases forms the genes that control all aspects of our heredity. As DNA molecules are incredibly long (a typical animal cell contains DNA with approximately 3x109 base pairs!) it is easy to see the vast number of combinations of bases that could exist, and the vast number of genes in each strand of DNA. Because the base pairs are always linked in the same way, (A with T, and C with G), if you were given only the order of the bases of one of the strands of DNA from a helix, you should be able to predict the bases and their order on the other, complementary strand.
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ACTIVITY 15 The principle of complementary base pairs in DNA (Allow about 5 minutes) As base pairing is a vital principle in understanding how DNA works, it is worth demonstrating your understanding of it before moving on. If a section of DNA has the following bases in the order given, what would the order and type of bases be on the other strand? Fill in the gaps you need only use letters to represent the bases.
DNA Backbone
Figure 43. One half of DNA base pairs
See Appendix 4 to check your base pair sequence. The principle of complementary base pairs means that one strand of DNA has the potential for being the template to create a new and complementary strand of DNA. This is known as replication and we will talk more about this later in this section. RNA A molecule related to DNA (deoxyribose nucleic acid) is ribose nucleic acid (RNA). This is involved in the transfer of information from DNA in the cell during the production of proteins (see transcription and translation later in this section). There is a small structural difference between RNA and DNA. As the name implies, RNA has ribose as the sugar in the backbone, not deoxyribose as in DNA. The difference is that ribose has an OH group instead of a hydrogen on one position on the pentose ring. Therefore deoxy means lack of an oxygen atom in comparison with the parent compound, ribose. The other key difference is that RNA has a different pyrimidine base that pairs with adenine (A). Uracil (U) takes the place of thymine (T). (See gure 44)
O H O N C C N R
Figure 44. Structure of uracil
C C
H H
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CHECKLIST You should now be able to: list the main parts of the DNA chain identify the two purine and two pyrimidine bases that are found in DNA state which bases pair together in the complementary base pairs describe the overall structure of DNA describe the structural differences between DNA and RNA.
6.3 Replication of DNA Before a cell divides into two, it must accurately duplicate its DNA so that the two new cells both carry the same genetic code. We have already looked at how one DNA strand can act as a template for a new strand. We are now going to consider this process, DNA replication, in more detail. DNA replication begins by the separation of the two complementary strands of a DNA double helix. Each of the strands is now available to act as a template for the formation of new DNA strands. The new DNA strands are built up by sequential addition of the deoxyribose nucleoside triphosphates. The correct deoxyribose nucleoside diphosphate is selected by the complementary base pair forming rst. Then the phosphodiester linkage is formed to clip the next unit on to the DNA chain. This process is repeated until each of the original DNA chains has a complete complementary DNA strand and two new double helices have been formed. Because of the presence in each double helix of one original strand and one new strand of DNA, the process is said to be semi-conservative (gure 45).
Original = DNA Strand New = DNA Strand
Figure 45. Semi-conservative process of DNA replication
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ACTIVITY 16 The semi-conservative process of DNA replication (Allow about 5 minutes) The semi-conservative process is important in the way new copies of DNA strands are produced. From the explanation so far, you should be able to say how the above sequence would continue. You can do this by completing gure 52 by drawing in the next four cells to show how many of those cells will contain original DNA from the rst single cell, and how many will contain only new DNA. You will nd the answer diagram in Appendix 4.
6.4 DNA transcription Once it had been determined that each specic protein consists of a unique sequence of amino acids (in the early 1950s), scientists speculated that this might be linked to the fact that DNA also has a linear sequence of sub-units. Proteins have a specic order of amino acids joined via peptide linkages, while DNA has a specic order of nucleotides linked via phosphodiester linkages. The relationship between DNA and proteins has since been worked out, and is discussed below. If you want to refresh your memory on proteins and their structure, refer back to the earlier section. DNA controls protein synthesis, but not directly. It uses a related compound called RNA to carry the appropriate messages to ensure that the right proteins with the right sequence of amino acids are made. However, before we look at the control of protein synthesis in detail, we need to understand the way that DNA transmits its messages that control protein manufacture. Copying into RNA The clue that suggested that DNA didnt control protein synthesis directly was that DNA is almost completely conned to the cell nucleus (eukaryotic cells) while it was known that protein is synthesised on the ribosomes which are found in the cell cytoplasm outside the nucleus. The RNA copies specic regions of the DNA chain, called genes. The RNA keeps the information of the DNA sequence that it copied, as well as the base pairing properties of DNA. Remember, though, that RNA uses the base uracil, where DNA would use the base thymine. Forming molecules of RNA from a DNA template is called DNA transcription. Like DNA replication, a strand of DNA acts as a template for the RNA ribonucleotides to match up against. Again, the RNA chain grows one ribonucleotide at a time by matching each new ribonucleotide up against the original DNA chain. In DNA replication, the original DNA helix comes apart and two new double helices of DNA are formed. In transcription, RNA is formed as a single chain. A single strand of DNA is also the template for RNA production. The DNA helix comes apart to provide a template for the formation of RNA, but just behind the point at which the ribonucleotides are being added to the strand of RNA, the DNA helix reforms and the RNA is released to move to where the protein synthesis will actually occur. Because RNA is only copying specic regions of DNA, the RNA molecules will be shorter than DNA. These short pieces of RNA direct protein synthesis and are called messenger RNA, or mRNA for short. There are other types of RNA that perform other functions, and we will explore that in more detail shortly.
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Translating the mRNA code Just as in Morse Code, where a series of dots and dashes and the order in which they are transmitted represent the different letters of the alphabet, in protein synthesis, groups of three nucleotides and the order in which they come in the mRNA chain represent amino acids. The process of turning this code into proteins is called translation. These groups of three nucleotides in the mRNA chain are called codons. Each group of three nucleotides represents (or codes for) a specic amino acid, and it isnt just the nucleotides that are present, but the order of the nucleotides which is critical in determining the amino acid being selected. With four different bases, there are 64 (43) possible combinations of the bases. These code for only 20 amino acids. This means that more than one codon or group of three bases can represent a given amino acid. For example, the following codons all represent alanine: GCU, GCC, GCA and GCG. Depending on where you start reading the RNA chain, i.e., starting with either the rst, second or third nucleotide in an RNA chain, there are three possible reading frames, and therefore three possible proteins that could be coded for, as shown in gure 46. However, usually, only one of the three possible reading frames results in a functional protein.
AUG
Met
GAU
Asp
GUA
Ala
UAC
Tyr
CG G
A AU
UGG
Trp
AUG
Met
UAU
Tyr
ACC
Thr
GGA
Gly
UGU
Cys
AUA
Ile
CCG
Pro
Figure 46. Reading frames on RNA chains
tRNA Just as DNA doesnt directly control protein synthesis, neither does mRNA. There is a nal link in the chain called transfer RNA, or tRNA. This allows the translation of mRNA into proteins. tRNA recognises the codons made up of three nucleotides on the mRNA, as well as recognising a specic amino acid. tRNA is made up of approximately 80 nucleotides which form a characteristic structure. This threedimensiona conformation is related to its function. This time, base pairs are formed between nucleotides on the same chain, and these interactions hold the tRNA chain in a shape which looks something like a clover leaf with three lobes. The amino acid attaches to one end of the chain (which always has a base sequence of -C-C-A). As was mentioned above, each type of tRNA will only bind one specic amino acid. Opposite where the amino acid is bound, there is a sequence of three nucleotides called the anticodon. These have bases complementary to those in the mRNA codon. Figure 47 shows schematically what happens during translation of mRNA (via tRNA) to proteins.
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mRNA C C A U C G C U A A A G C G U G G A G C A
U C C
Pro
C C A
C C A
Ser
Leu
Lys
Arg
Gly
Incoming tRNA with amino acid attached
Growing polypeptide chain

Figure 47. Translation of mRNA via tRNA to proteins
The nal part of the jigsaw that completes the set of structures necessary for translation is the ribosome. This is where the process in gure 48 takes place. Ribosomes are a combination of more than 50 proteins along with some structural RNA (rRNA). The ribosome reads the code provided by the mRNA. It rst nds the specic start site on the mRNA that will set the reading frame. This in turn species the amino acid which will be at the amino terminus of the protein to be formed. The ribosome then moves along the mRNA molecule reading the codons one by one. As each codon is read, a tRNA molecule with the appropriate anticodon that matches the codon moves into place, and the amino acid it is carrying is added to the growing protein chain. Eventually, the end of the mRNA code is reached, and both the ribosome and the end of the protein which was attached to the ribosome (the carboxyl terminus) are released.
Ribosome subunits released mRNA
Ribosome
Start Stop Growing polypeptide Complete polypeptide released

Figure 48. Ribosomes involved in the production of proteins
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ACTIVITY 17 Processes involving DNA and RNA (Allow about 10 minutes) You have now learned about three processes involving one or both of DNA and RNA. To assess your understanding of which is involved where, look at each process in turn. Write down the products obtained at each stage (e.g., DNA, protein etc.), and the molecules (DNA, mRNA, etc.) involved. To check your answers, see Appendix 4.
Process Replication Molecules involved Products
Transcription
Translation
6.5 Summary DNA is fundamental in both passing on genetic material and in the production of proteins in living things. It is made up of nucleotide sub-units, which form long chains linked by phosphodiester linkages. The chains of nucleotides coil together in pairs to form the DNA double helix with the sugar-phosphate backbone on the outside and complementary base pairs on the inside. DNA produces more DNA via a semi-conservative process called replication and it also provides a template for mRNA production via a process called transcription. mRNA consists of codons, groups of three nucleotides which represent a specic amino acid. These codons direct the formation of proteins via tRNA which delivers the individual amino acids to be joined together to form a protein via a process called translation. Ribosomes, which consist of a combination of proteins and rRNA, are where this process takes place.
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CHECKLIST You should now be able to: explain the process of replication, including the addition of nucleotides to a DNA chain and the principle of semi-conservative DNA replication derive the base order for a new strand of DNA given the original DNA sequence explain the process of DNA transcription _ explain the process of translation including the structure and role of codons explain the role of tRNA and ribosomes in translation.
References Proteins 1. C.H. Wynn, The Structure and Function of Enzymes, 2nd ed, Edward Arnold, 1979. 2. L. Stryer, Bio Chemistry, 2nd ed, W.H. Freeman and Company (San Francisco), 1981. 3. A. Lehninger, Principles of Bio Chemistry, Worth Publishers Inc., 1982. 4. B. Alberts et al., Molecular Biology of the Cell, 2nd ed, Garland Publishing Inc. (New York and London), 1989. Enzymes 1. C.H. Wynn, The Structure and Function of Enzymes, 2nd ed, Edward Arnold, 1979. 2. D. Freifelder, Molecular Biology, 2nd ed, Jones & Bartlett Publishers Inc. (Boston), 1987. 3. L. Stryer, Bio Chemistry, 2nd ed, W.H. Freeman and Company (San Francisco), 1981. 4. A. Lehninger, Principles of Bio Chemistry, Worth Publishers Inc., 1982. Genetics and Cell Replication 1. B. Alberts et al., Molecular Biology of the Cell, 2nd ed, Garland Publishing Inc. (New York and London), 1989. 2. A. Lehninger, Principles of Bio Chemistry, Worth Publishers Inc., 1982. 3. L. Stryer, Bio Chemistry, 2nd ed, W.H. Freeman and Company (San Francisco), 1981.
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APPENDIX 2 ACTIVITY 13 Optical isomers of amino acids The diagram you produced should look like this.
H2N
COOH
HOOC
NH 2
mirror image
ACTIVITY 14 Inuence of amino acid side chains 1. A 2. C 3. D 4. E 5. B
ACTIVITY 15 Formation of a tripeptide The condensation could take place in two ways to give two possible answers.
O O H2N C O H N CH3 N H OH C O CH3 H2N N H C O H N
O C OH
Condensation at the left hand end of the dipeptide
Condensation at the right hand end of the dipeptide
The structure of the bonds linking the amino acids should look the same as the ones linking the two amino acids of the dipeptides. ACTIVITY 16 Levels of protein structure Level of structure Bonds and/or forces involved Primary Secondary Tertiary Peptide bonds, i.e., covalent bonds Hydrogen bonds between components of the peptide bond Covalent bonds, hydrogen bonds, ionic forces, Van der Waals forces, hydrophobic interactions between different parts of one peptide chain As for tertiary, except the interactions are between different polypeptide chains
Quaternary
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BIO CHEMISTRY
APPENDIX 3 ACTIVITY 17 Classication of enzymes 1. C 2. F 3. A 4. E
5. B
6. D
ACTIVITY 18 Enzyme structure and mechanism A) You could have mentioned covalent, ionic and hydrogen bonds, Van der Waals forces, steric interactions. B) (A) is lock and key. The substrate will slot into the complementary active site in the enzyme like a key into a lock! (B) is induced t. This is like the lysosyme example where the enzyme changes shape as it binds the substrate, so its active site only completely ts the substrate once the substrate is bound. ACTIVITY 19 Effect of different inhibitors Your graphs should have looked like this:
Vmax Vmax
x Time
x Time
For both cases, addition of the inhibitor causes the enzyme to stop processing the substrate. In competitive inhibition, addition of a large excess of substrate will counteract the inhibitors effect, but for non-competitive inhibition, this will have no effect. In this case, the enzyme catalysed reaction will only begin again once the inhibitor has been removed.
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APPENDIX 4 ACTIVITY 20 Basic structure of DNA
O O
-
P O O A CH 2 O
O OP O O C H2 O T
O OP O O C H2 O C
O O
-
P O O G CH 2 O
This is the kind of structure you should have drawn. As I didnt specify the order that the nucleotides should be in, dont worry if the order is different from what is shown here.
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ACTIVITY 21 The principle of complementary base pairs in DNA
A T
C G
C G
G C
T A
A T
ACTIVITY 22 The semi-conservative process of DNA replication The two original DNA strands are again taken through into the new cells, but two of the new cells have entirely new DNA in comparison with the original cell.
ACTIVITY 23 Processes involving DNA and RNA Process Replication Molecules involved Products
DNA strands, DNA sub-units More DNA a complementary strand to the template DNA strands, mRNA strands, mRNA RNA sub-units mRNA, tRNA, rRNA (in ribosomes), amino acids Protein
Transcription Translation
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BIO CHEMISTRY

Module 1 Unit 5 Bio Chemistry

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To move through this unit, select the buttons shown on the screen that look like this.

Module 1 Unit 5 Bio Chemistry

Society of Cosmetic Scientists Distance Learning

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

There are several possibilities. I have reproduced two in gure 1.

Figure 1. Possible arrangements of 3C: 6H: 3O

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

CHO Unspecific representation (no information on structure) HO C H HO C H HO C H H C OH CH 2OH

Glucose: an aldo-hexose Carbon atom number H C O H C OH HO C H H C OH H C OH H C OH H 1 2 3 4 5 6

Figure 2. Structure of hexose sugars

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

CHO HCOH HOCH HCOH HCOH H2COH

HCOH HCOH HOCH HCOH HC H2COH O

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

1.4 More complex carbohydrates

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

Figure 5. Condensed sugars

Figure 6. Alternative possibility for linking glucose molecules

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

Figure 7. Branching arrangement of polysaccharide chains

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

Figure 10. Structure of a triacylglycerol

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

2.4 The implications of unsaturation

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

Figure 11. The shapes of double bonds in hydrocarbon chains

Figure 12 cis unsaturated hydrocarbon chains preventing close packing

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

polar/hydrophilic will mix with water

nonpolar/hydrophobic will not mix with water

Figure 13. Structure of a phospholipid

CH3 CH3 CH2 CH2 CH2 CH2 CH HO CH3 C C CH2 CH CH CH CH2 C CH CH CH

CH2 CH2 CH2 CH2 CH CH3 CH3

Figure 14. Structure of a typical steroid

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

R CH amino group H2N

side chain group COOH carboxylic acid group

DIPLOMA/CERTIFICATE IN COSMETIC SCIENCE MODULE 1 UNIT 5

Figure 18. Ionisation of amino acids

General structure of amino acids Basic (polar, positively charged*) Non-polar

NH2 Lysine (E)

Isoleucine (E) Leucine (E)

His Phenylalanine (E) Phe

Acidic (polar, negatively charged*)

Aspartic acid (Aspartate) Glutamic acid (Glutamate)

Glu Tryptophan (E)

Neutral (polar, non-charged*)

Ser Tyrosine Tyr

CONH2 CH2 CONH2 CH2 CH2

Special amino acids Asparagine Asn