BIOCHEMISTRY

VINAYAKUMAR ROHIT

SECTION:B

ID NO:-17-0169-909

1.How is enzyme activity regulate via the following mechanisms:

A) COVALENT MODIFICATION:

The activity of enzyme can be increased or decreased by covalent modification .It means either
addition of a group to the enzyme protein by a covalent bond or removal of a group by cleaving a
covalent bond. This is a reversible reaction. The most common type of covalent modification is the
reversible protein phosphorylation. This means covalent attachment of one or several phosphate
groups to the protein

ADP P ADP

Phosphorylation

(kinase)

PROTEIN PROTEIN P

Dephosphorylation

(phosphatase)

SCHEMA OF COVALENT MODIFICATION

A protein kinase moves a phosphate group(p) from (ADP(p)) to the protein. The biological properties
of the protein is thereby altered. There is also a protein phosphatise that is able to remove the
phosphate group. The amount of phosphate that is associated with the protein is thus determined
by the relative activation of the kinase and the phosphatase

B) GENE REGULATION:

Regulation of gene expression includes a wide range of mechanisms that are used by cells to
increase or decrease the production of specific gene products (protein or RNA), and is informally
termed gene regulation. Gene expression can be regulated by various cellular processes with the
aim to control the amount and nature of the expressed genes. These regulatory proteins bind to
DNA and send signals that indirectly control the rate of gene expression. Most mechanisms
that control gene expression do so by controlling transcription, the synthesis of mRNA. However
there are other mechanisms for controlling the rate of protein synthesis that occur downstream
between transcription and translation.

2.Explain the difference between competitive, Non-competitive, and uncompetitive

enzyme inhibition.

1.COMPETITIVE INHIBITION:-

In competitive inhibition, the inhibitor competes with substrate for binding to the active site. When
the inhibitor occupies the active site, it forms an enzyme-inhibitor complex and the enzyme cannot
react until the inhibitor dissociates. Such inhibitors are substrate analogs,since they have a structure
similar to the substrate but are unreactive.
SCHEMA OF COMPETATIVE INHIBITION
EG:-Antintoplastic drug methotrexate is an example of competitive inhibitor. Methotrexate has a
structure similar to that of the vitamin folic acid. It acts by inhibiting the enzyme dihydrofolate
reductase ,preventing the regeneration of dihydrofolate from tetrahydrofolate. This interfers with
DNA synthesis and blocks cell division in rapidly dividing cancer cells.

2.NON-COMPETITIVE INHIBITION:-

In non-competitive inhibition,the non competitive inhibitor bind to a site other than the active site
known as the allosteric site on the enzyme.In this case inhibitors are usually structurally unrelated to
the substrate.Thus both the substrate and inhibitor theoretically can bind to the enzyme at the same
time.The inhibitor usually disorts the active sites,thus altering conformations of their catalytic
residues when further reduces the effectiveness of the enzyme substrate complex.In this case the
effect of non competitive inhibitor cannot be reversed by increasing substrate concentration.

SCHEMA OF NONCOMPETATIVE INHIBITION

EG:-Strychnine is a colourless highly toxic alkaloid that causes muscular convulsions and eventually
death throghh asphyxia. Strychnine binds to glycine receptors at allosteric site preventing glycine
from binding.This causesmotor neuron to continuously fire and the victim has constant muscle
contraction.

3.UNCOMPETITIVE INHIBITION:-

In an uncompetitive inhibition , the inhibitor only binds to the enzyme-substrate complex.The

formation of its binding site is only formed after the enzyme and the substrate has been intracted
among themselves.The uncompetitive inhibitor does not work when additional substrates are trying
to be involved.The enzyme-substrate inhibitor complex does not produce any product.

SCHEMA OF UN COMPETATIVE INHIBITION

3.What are the different mechanism of enzyme action?

There are mainly two theories which explain the mechanisms and enzyme action:

a)Fischer’s Lock and Key Theory:-

According to this theory first a physical contact is made between the enzyme and the substrate.As
only a specific key fits in a particular lock to open it ,a specific substrate combines with the active site
of a specific enzyme as a result of which an enzyme-substrate complex is formed.Then the enzyme
acts on the substrate to produce products. After the reaction enzyme is released from the enzyme-
substrate complex and is ready to bind with other substrate.When a dissimilar substrate approaches
the enzyme, it cannot combine with the active site of the enzyme.

ENZYME SUBSTRATE

b)Koshland’s induced Fit Theory:-

It was proposed by Koshland. According to this theory the substrate induces the enzyme to adjust its
shape leading to the formation of enzyme-substrate complex as proteins are not rigid. Then enzyme
acts on substrate to produce products. The substrate –active site are flexible but structurally not
complementry .The binding of a substrate to the active site is believed to induce a slight alteration in
the shape of active site .After the release of products the active site return to its original
configuration .

ENZYME SUBSTRATE

4.How are nucleotides synthesized and metabolized? Indicate possible disease conditions if there is
a block in the metabolic pathway.

Nucleotides can be separated into purines and pyramidines .They are both primarily produced in the
liver .The both contain sugar and phosphate, but have nitrogenous bases that are different sizes.
However, all nucleotide synthesis requires the use of phosphoribosylpyrophospate {PRPP} which
donates the ribose and phosphate necessary to create a nucleiotide .Pyrimidine nucleotides include
cytidine, uridine,and thymidine. The synthesis of any pyrmidine nucleotide begins with the formation
of uridine .The reaction requires aspartate, glutamine, bicarbonate, and 2 ATP molecules.

Nucleotide metabolized in the process by which nucleic acid are synthesized and degraded. Nucleic
acids are polymers of nucleotides. Nucleotide is an anabolic metabolism generally involving the
chemical reaction of phosphate .Nucleobasis can be salvaged to recreate new nucleotide .other
synthesis and degradation reactions require enzyme to falitate the event.
Defects and deficiencies can lead to LESCH –NYHAN SYNDROME is caused by deficiency in
hypoxanthine-guanine phosphoribosyltransferase or HGPRT, The enzyme that catalyzes the reversible
reaction of producing guanine from GMP. This is a sex linked congenital defect that causes
overproduction of uric acid along with mental retardation, spasticity , and a urge to self mutilate .

5. Create a summary of the metabolic processes of glycolysis ,citric acid cycle ,electron transport chain
,beta oxidation of fats and protein metabolism in a single page.

GLYCOLISIS :-

For simple fermentations, the metabolism of one molecule of glucose to two molecules of pyruvate
has a net yield of two molecules of ATP. ... Cells performing aerobic respiration synthesize much more
ATP, but not as part of glycolisis. These further aerobic reactions use pyruvate and NADH + H from+

glycolysis.

CITRIC ACID CYCLE:-

The total energy gained from the complete breakdown of one (six-carbon) molecule of glucose by
glycolysis, the formation of 2 acetyl-CoA molecules, their catabolism in the citric acid cycle, and
oxidative phosphorylation equals about 30 ATP molecules, in eukaryotes.

ELECTRON CHAIN TRANSPORT:-

chain of the thylakoid membrane. The electron transport chain in the mitochondrion is the site of
oxidative phosphorylation in eukaryotes.

The NADH and succinate generated in the citric acid cycle are oxidized, providing energy to power ATP

synthase. Photosynthetic electron transport chain of the thylakoid membrane.

BETA –OXIDATION OF FAT AND PROTEIN METABOLISM:-

Beta-oxidation is the catabolic process by which fatty acid molecules are broken down in the cytosol
in prokaryotes and in the mitochondria in eukaryotes to generate acetyl-CoA, which enters the citric
acid cycle, and NADH and FADH , which are co-enzymes used in the electron transport.
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