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LSB231- Physiology:

Lecture 1.

Part A: The Cell


organelles
cytoskeleton
k l
www.invitrogen.com
LSB231- Physiology:
Lecture 1.

Part B: The Plasma Membrane


membrane structure
membrane p
proteins
membrane transport
www.smbs.buffalo.edu/bch/Labs/Kosman/research.htm
LSB231- Physiology:
Lecture 1.
Part C: Metabolism and Energy
types of metabolism
role of ATP
sources of ATP

www.invitrogen.com
LSB231- Physiology:
Lecture 1.
• Part A: The Cell
– organelles
g
– cytoskeleton
• Part B: The Plasma Membrane
– membrane structure
– membrane proteins
– membrane transport
• Part C: Metabolism and Energy
– types of metabolism
– role
l off ATP
– sources of ATP
LSB231- Physiology:
Lecture 1.
Learning Objectives – Part A
• Identify each of the major cellular organelles

• Describe
D ib the
th ffunction
ti off each
h organelle
ll

• Identify the major intracellular structural proteins


which make up p the various cytoskeletal
y structures

• Describe the function of each of the major


intracellular cytoskeletal structures
Part A: The Cell
• Smallest living
component of the
human body.
body

Organ Tissue Cells


The Cell
• Smallest living
component of the
human body.
body
• Bounded by plasma
membrane.
b
The Cell
• Smallest living
component of the
human body.
body
• Bounded by plasma
membrane.
b
• Intracellular fluid is
called the
cytoplasm.
• Contains functional
subunits called
organelles.
Organelles

• Membrane bound.
• Analogous to organs
found within the
body.
p
• Each serve specific
functions.
• Supported by
cytoskeleton.
Organelles

• Nucleus
• Endoplasmic
reticulum
• Golgi apparatus
• Mitochondria
• Lysosomes
• Peroxisomes
P i
The Nucleus

• Bounded by nuclear
envelope
l with
ith pores.
The Nucleus

• Bounded by nuclear
envelope
l with
ith pores.
• Contains DNA in the
form of chromatin.
The Nucleus

• Bounded by nuclear
envelope
l with
ith pores.
• Contains DNA in the
form of chromatin.
• Mayy display
p y
nucleolus.
The Nucleus

• Bounded by nuclear
envelope
l with
ith pores.
• Contains DNA in the
form of chromatin.
• Mayy display
p y
mRNA
RNA
nucleolus.
• Site of DNA synthesis.
• Site of transcription.
Endoplasmic Reticulum
• Outer membrane
continuous with
nuclear envelope.
Endoplasmic Reticulum

• Outer membrane
continuous
ti with
ith
nuclear envelope.
• May be characterised
as either
– “smooth” or
– “rough”
• depending upon
presence of
p
ribosomes.
Endoplasmic Reticulum
• Rough
g ER (RER)
( ) tRNA mRNA
– site of translation for
proteins that are aa
transported out of the
cell or into other
organelles.
ll Peptides
– Synthesis of phospholipid.
Endoplasmic Reticulum
• Smooth ER (SER)
– lipid metabolism
– cholesterol/steroid
h l l id
synthesis
– drug
d g metabolism
t b li (li
(liver))
– calcium storage
(eg sarcoplasmic reticulum
of skeletal muscle cells)

Ca2+
Golgi Apparatus
• Receives packages of
proteins from RER.
• Modifies proteins:
– p
post-translational
modifications.
– Glycosylation.
• Sorts and packages
p ote s:
proteins:
– vesicles delivered to other
g
organelles or p
plasma
membrane.
Mitochondria

• Produce energy source:


– Adenosine Triphosphate
(ATP).
Mitochondria
Outer membrane

• Produce energy source:


– Adenosine Triphosphate
(ATP).
• Consist of inner and outer
membranes, surrounding a
fluid matrix.

Matrix

Inner membrane
Mitochondria

• Produce energy source:


– Adenosine Triphosphate
(ATP).
• Consist or inner and
d outer
membranes, surrounding a
fl id matrix.
fluid i
• Contain mitochondrial DNA
– mitochondrial disorders may
therefore be maternally
inherited.
Lysosomes

• Bound by single
lysosomes
membrane.
b
• Contain acidic
environment.
g
• Serve digestive function.
• Contain acid hydrolases.
– Proteases,
Proteases lipases etc
etc.
• Especially abundant in
phagocytic cells.
cells Neutrophil leukocyte
– eg. Neutrophils
Peroxisomes

• Bound by single
membrane.
b
• Contain enzymes required
for metabolism of toxins.
• Peroxisomal Disorders:
– Can cause serious
developmental and
neurological defects.
– eg. X-linked Peroxisomes in liver cell
adrenoleukodystrophy
Cytoskeleton
• Internal scaffold of cells.
• Controls cell shape.
• Controls cell transport.
• C i t of:
Consists f
– microfilaments (actin
polymer)
– microtubules (tubulin
polymer)
– intermediate filaments
(vary in composition: eg.
eg Actin filaments
Keratins). in fibroblasts
Microfilaments
G-actin
G actin (monomer)
• Composed of
polymerised actin.

F-actin (polymer)
Microfilaments
• Composed of or = G-actin (monomer)
polymerised actin.
• Associated with
numerous actin-
binding proteins, eg. F-actin (polymer)
myosin.
Microfilaments
• C
Composed d off
polymerised actin.
• Associated with
numerous actin-
binding proteins, eg.
myosin.
•Human Neutrophil seeks
• Essential for bacteria
regulating
g g changes
g in for permanent
•for
cell shape. relationship
•Enjoys food and long
– Amoeboid motilityy amoeboid crawls around
– Muscle contraction. the body
Microfilaments
• Composed of
polymerised actin.
• Associated with
numerous actin-
binding proteins, eg.
myosin.
• Essential for
regulating changes in
cell shape. Movie of Leukocytes hunting
bacteria
– Amoeboid motility ...if
if you don’t
don t come to the
– Muscle contraction. lecture you don’t get to see
the movie!!
Microtubules

• Composed of
polymerised
l i d ttubulin.
b li
Microtubules

• Composed of
polymerised
l i d ttubulin.
b li
• Associated with motor
proteins eg. kinesin.
Microtubules

• Composed of
polymerised
l i d ttubulin.
b li
• Associated with motor
proteins eg. kinesin.
pp
• Support cell structure.
• Provide paths for
intracellular
transport.
– Eg.
Eg Neuro
Neuro-transmitters
transmitters.
Intermediate Filaments

• Composition varies
according
di to
t cell
ll ttype.
• Less dynamic than
microfilaments and
microtubules.
• “Tough” insoluble
fibres.
Intermediate Filaments

• Composition varies
according
di to
t cell
ll ttype.
• Less dynamic than
microfilaments and
microtubules.
• “Tough” insoluble
fibres.
• Example:
– keratins in epithelial
cells.
Intermediate Filaments

• Composition varies
according
di to
t cell
ll ttype.
• Less dynamic than
microfilaments and
microtubules.
• “Tough” insoluble
fibres.
• Example: Human corneal epithelial
cells stained with antibody
– keratins in epithelial
to keratin 3
cells.
Part B: The Plasma Membrane
Learning Objectives – Part B
• Describe the structure and identify the various components of the
plasma membrane

• Describe the general roles of transmembrane or integral proteins

• Describe the processes of membrane transport, diffusion and osmosis

• Understand
d d and
dddescribe
b osmolarity
l

• Describe and understand the difference between facilitated diffusion


and
d active
i transport

• Describe the fluid composition of some common ions inside and


outside
id the
h cellll
Part B: The Plasma Membrane

• Double layer of
phospholipid
h h li id and d
cholesterol.

Polar
Non-polar
Membrane Structure

• Double layer of
phospholipid
h h li id and d
cholesterol.
Membrane Structure
Crystal of lipophilic dye
• Double layer of
phospholipid
h h li id and d
cholesterol.
• Highly dynamic.
Fluid Mosaic Model

• Double layer of
phospholipid
h h li id and d
cholesterol.
• Highly dynamic.
• Accessoryy
molecules:
– proteins
p
• integral
• peripheral
– carbohydrates
Membrane Proteins

• Roles of membrane
proteins:
t i
• Structural support.
• Communication.
• Channels.
• Gates.
• Pumps.
Transmembrane protein
Membrane Proteins

• Communication.
• Provide receptors for
soluble ligands.
• eg.
g hormones.
h
Membrane Proteins

• Communication.
– Provide receptors for
soluble ligands.
• Provide
P id receptorst ffor
other cells.
Membrane Proteins

• Communication.
– Provide receptors for
soluble ligands.
– Provide
P id receptorst ffor
other cells.
– Provide channels
between cells.
• Gap Junctions
between muscle cells.
Membrane Transport

• The plasma membrane is a semi-permeable


b i
barrier.
Membrane Transport

• The plasma membrane is a semi-permeable


b i
barrier.
• Transport of molecules across membrane is
dependent upon:
– relative solubility.
– physical size.
– presence of concentration gradient.
– other factors: specialised transport proteins.
Membrane Transport
• Solubility:

– more hydrophobic/more non-polar


molecules readilyy dissolve through
g lipid
p
bilayer.
Membrane Transport
• Solubility:

– more hydrophobic/more non-polar


molecules readilyy dissolve through
g lipid
p
bilayer.

– More hydrophilic/polar molecules have


poor solubility in lipid bilayer.
Membrane Transport

• Size:

– certain small molecules can pass through


protein channels in the plasma membrane:
– examples:
• Aquaporins: water channels.
channels
• Ion channels: tend to be specialised for
each ion.
– may be “gated”: open in response to stimulus.
Membrane Transport

• Basic channels:
– Always open.
– Enable passive
t
transportt off
molecules across
plasma membrane by
diffusion.
– Restricted to
molecules of certain
physico-chemical
properties.
Membrane Transport

• Gated channels:
– Normally closed.
Membrane Transport

• Gated channels:
– Normally closed.
– Open in response to
appropriate
i t stimulus.
ti l
• Hormone
• Neurotransmitter
• Ion balance (voltage)
Diffusion

• Diffusion:
– movement of
substance along a
concentration
gradient.
– Results from random
collisions of
particles known as
p
Brownian Motion.
Osmosis

• Diffusion:
– movement of
substance along a
concentration
gradient.
• Osmosis:
– diffusion of water
along a concentration
gradient.
Osmosis

• Diffusion:
– movement of
substance along a
concentration
gradient.
• Osmosis:
– diffusion of water
along a concentration
gradient.
Osmosis

• Diffusion:
– movement of
substance along a
concentration
gradient.
• Osmosis:
– diffusion of water
along a concentration
gradient.
– Causes increase in
hydrostatic pressure
within the cell.
Osmolarity

• Direction of osmosis is determined by relative


osmolarity:
l it
– total number of solute particles per litre of
solution.
l ti

• Sometimes use term osmolality:


– total number of solute particles per litre of water.
Osmolarity

• Example: for a 0.15 molar solution of


N Cl the
NaCl th osmolarity
l it iis d
double:
bl iie 0
0.3
3
mosmol.
Na
Na
Cl
Dissociates
into two
Cl ti l
particles
Osmolarity

• Osmosis occurs when intra-cellular osmolarity is


different to the external osmolarity.
osmolarity
• Typical osmolarity within cells is: 300 mosmol.
Osmolarity

• Osmosis occurs when intra-cellular osmolarity is


different to the external osmolarity.
osmolarity
• Typical osmolarity within cells is: 300 mosmol.

• 300 mosmol solutions are iso-osmotic/iso-tonic.


• >300 mosmol solutions are hyper-osmotic/hyper-
tonic.
• <300 mosmol solutions are hypo-osmotic/hypo-
tonic.
tonic
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
concentration
gradient.
gradient
– Involves binding of
molecule to
transporter protein.
Facilitated Diffusion

• Facilitated diffusion:
– requires
i
concentration
g
gradient.
– Involves binding of
molecule to
t
transporter
t protein.
t i
– Example: Glucose
transport by GLUT
proteins (insulin
increases levels of
GLUT4).
GLUT4)
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy.
• ATP
• Ion gradient
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy.
• ATP
• Ion gradient
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy.
ATP
– Example with ATP.
ATP
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy. P
– Energy released from ADP
phosphate bond.
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy. P
– ADP released.
released

ADP
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy
entropy.
– Requires energy. P
– Energy used to alter
orientation of
protein.
p ADP
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy
entropy.
– Requires energy. P
– Molecules released.
released

ADP
Active Transport

• Pumps:
– Useful for producing
concentration
gradients.
gradients
– Working against
entropy.
entropy
– Requires energy.
– Phosphate released.
released
– Protein returns to
original ADP
conformation. P
Active Transport

• Pumps: • Calcium-ATPase
– Useful for producing – maintains low intracellular
concentration calcium concentration.
gradients
gradients.
• Sodium/potassium-
– Working against
ATPase
entropy
entropy.
– pumps 3 sodium ions out
– Requires energy. for every 2 potassium ions
– Examples: in.
Fluid Composition
Outside cells Inside Cell
• Na+ 140 mM 15 mM

• K+ 4 mM 150 mM

• Cl- 110 mM 10 mM

• Ca2+ 1mM 0 001 mM


0.001
Part C: Metabolism and Energy
Learning Objectives – Part C

• Define Metabolism and Energy

• Describe the various pathways which cells use to


produce ATP ie Glycolysis
Glycolysis, the Krebs cycle
cycle, and
the Electron Transport Chain

• Describe the process of ATP production by


Creatine Kinase in muscle cells
Part C: Metabolism and Energy

• Metabolism:
– refers to the collection of chemical reactions
that take place within a living structure.
Part C: Metabolism and Energy

• Metabolism:
– refers to the collection of chemical reactions
that take place within a living structure.
• Catabolic reactions:
– involve destruction of molecules.
Part C: Metabolism and Energy

• Metabolism:
– refers to the collection of chemical reactions
that take place within a living structure.
• Catabolic reactions:
– involve destruction of molecules.
• Anabolic
A b li reactions:
ti
– involve construction of molecules.
Energy
• Energy can neither be created nor
destroyed it simply changes form:
– First Law of Thermodynamics.
Energy
• Energy can neither be created nor
destroyed it simply changes form:
– First Law of Thermodynamics.
• Cells acquire energy via catabolism of
carbohydrates, fats and proteins.
Energy
• Energy can neither be created nor
destroyed it simply changes form:
– First Law of Thermodynamics.
• Cells acquire energy via catabolism of
carbohydrates, fats and proteins.
• Most of this energy is lost as heat.
heat
Energy
• Energy can neither be created nor
destroyed it simply changes form:
– First Law of Thermodynamics.
• Cells acquire energy via catabolism of
carbohydrates, fats and proteins.
• Most of this energy is lost as heat.
heat
• Remainder is stored in the form of
ATP.
ATP
Adenosine Triphosphate

• Energy is stored within phosphate


bonds.
bonds
• Analogous to potential energy stored
when
h a springi iis compressed.
d
Sources of ATP
Carbohydrates Cytoplasm
Glycolysis
Muscle
C ti
Creatine
Pyruvate Lactate

Fats/Proteins
ATP
Krebs Cycle

ADP
Fats NADH + H+ and FADH2
Creatine-P
Oxygen Electron Transport
Chain
Mitochondria
Glycolysis

• Conversion of glucose to pyruvate.


• Occurs within the cytoplasm.
cytoplasm
• Can occur in absence of oxygen
(
(anaerobic).
bi )
• Net yield: 2 molecules of ATP.
• 1 gram of glucose yields 4 kcal.
Glycolysis

• Conversion of glucose to pyruvate.


• Occurs within the cytoplasm.
cytoplasm
• Can occur in absence of oxygen
(
(anaerobic).
bi )
• Net yield: 2 molecules of ATP.
• 1 gram of glucose yields 4 kcal.

• 1 calorie = energy required to increase


the temperature of 1 gram of water by
1º C.
Glycolysis

• Conversion of glucose to pyruvate.


• Occurs within the cytoplasm.
cytoplasm
• Can occur in absence of oxygen
(
(anaerobic).
bi )
• Net yield: 2 molecules of ATP.
• 1 gram of glucose yields 4 kcal.

• Also produces 2 (NADH + H+)


– utilised
tili d dduring
i g Electron
El t T
Transportt Ch
Chain.
i
Krebs Cycle
• Occurs within mitochondrial matrix.
• Pyruvate first converted to Acetyl-CoA.
• Acetyl-CoA
Acetyl CoA enables cycling of reactions between
citrate and oxaloacetate (Citric Acid Cycle).
• Requires aerobic conditions to proceed.
• Net yield for each cycle: 1 molecule of ATP.

Matrix
Krebs Cycle
• Occurs within mitochondrial matrix.
• Pyruvate first converted to Acetyl-CoA.
• Acetyl-CoA
Acetyl CoA enables cycling of reactions between
citrate and oxaloacetate (Citric Acid Cycle).
• Requires aerobic conditions to proceed.
• Net yield for each cycle: 1 molecule of ATP.

• Also produces 3 (NADH + H+) and 2 FADH2


– utilised during Electron Transport Chain.
Krebs Cycle
• Acetyl-CoA
Acetyl CoA can also be generated from:
– protein: in the form of amino acids (via
pyruvate)
– Fats: in the form of fatty acids.
Krebs Cycle
• Acetyl-CoA
Acetyl CoA can also be generated from:
– protein: in the form of amino acids (via
pyruvate)
– Fats: in the form of fatty acids.
• E
Each
h gram off amino
i acids
id produces:
d 4kkcall
• Each gram of fatty acid produces: 9 kcal
Electron Transport Chain
Outer membrane
• Occurs across inner
mitochondrial
membrane.
• Oxygen is
essential.
• Utilises:
U ili
– FADH2
– NADH + H+ Matrix

Inner membrane
Electron Transport Chain

• Hydrogen ions from FADH2 and NADH + H+


are deposited into the fluid matrix.
matrix
• Produces a concentration gradient of H+
ions.
ions
• ATP is generated as the H+ ions flow back
across the
h iinner membrane
b via
i an ATP
ATP-
synthase.
Electron Transport Chain

• Net yield:
– 3 molecules of ATP from each NADH + H+.
– 2 molecules of ATP from each FADH2.
Creatinine Phosphate

• Muscle cells have an


additional method
for producing ATP.
Creatine Phosphate
Creatine Creatine Pi
• Muscle cells have an
additional method
Creatine Kinase
for producing ATP.

Glycolysis
Krebs Cycle ATP + Pi
ADP
ETC

Muscle Contraction/Relaxation
See you on Monday morning for your first Prac
But for now…
now

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