Metabolism 1: An Introduction to Protein Structure
Outline the reaction by which amino acids are joined together; sketch a trimeric peptide, illustrating
the amino terminus, carboxyl terminus and side chains
Protein: any of a group of organic compounds composed of one or more chains of amino acids and
forming an essential part of all living organisms

Human body: 20% protein

Protein mutations are responsible for a variety of diseases (e.g. sickle cell anaemia)
Structure of an amino acid:

Substitutions of the side chain (R group) give rise to the 20 different amino acids

Backbone: the whole of the amino acid minus the side chain
Amino acids with hydrophobic side chains
Amino acids with hydrophilic side chains
Glycine: the simplest amino acid (R group = H)
Asparagine
Alanine
Glutamine
Valine
Cysteine
Leucine
Histidine
Isoleucine
Serine
Proline
Threonine
Methionine
Tyrosine
Phenylalanine
Tryptophan
Aspartate
Glutamate
Lysine
Arginine
Amino acids with charged side chains
Arginine and Lysine
Protonated at physiological pH; therefore basic
Histidine
Often protonated
Aspartic acid and Glutamic acid
Deprotonated at physiological pH; therefore acidic
Amino acids with charged side chains:

The ionisation state of an amino acid provides vital biological properties to many proteins

Therefore cells cannot generally tolerate wide changes in pH

If the ionisation state of key amino acids within a protein is altered: a loss of biological activity often
results

The ability to take up and release protons gives amino acids some buffering capacity to resist some
changes in pH
Chirality:

The central Cα carbon atom is a chiral centre: i.e. it has 4 different substituents bound to it

This produces optical isomers (enantiomers) of each amino acid, which are non-superimposable mirror
images of each other

Glycine (Gly) has no side chain; therefore it is the only non-chiral amino acid

All amino acids found in proteins are L-enantiomers
Peptides are formed by a condensation reaction between 2 amino acids:
Structure of a peptide:
Protein structure:

The polypeptide chain of a protein rarely forms a disordered structure

Proteins have functions which rely upon specificity

Functionality requires a definite 3D structure (conformation) of the polypeptide chain

Proteins generally possess a degree of flexibility which is necessary for function (e.g. muscle fibres)
Characteristics of the peptide bond:

There is no free rotation about the peptide bond

The C=O and N–H groups are in the same plane of the molecule

The other 2 bonds in the backbone of the polypeptide chain are able to rotate
Understand the concepts of primary structure, secondary structure, tertiary structure & quaternary
structure with respect to proteins
Folding of proteins:

Proteins generally fold into a single conformation of lowest energy

Folding of proteins may occur spontaneously or it may involve chaperones
Chaperones: molecules which bind to a partially folded polypeptide chain and ensure that folding
continues along the most energetically favourable pathway

By breaking the bonds which hold a protein together, a protein is denatured into the original flexible
polypeptide
Common laboratory denaturants include:
o
Urea: breaks hydrogen bonds
o
2-mercaptoethanol: breaks disulphide bonds
Structural levels of proteins:

Primary structure: the linear sequence of amino acids which constitute the protein
Nomenclature: the protein sequence is written from the amino terminus to the carboxyl terminus

Secondary structure: local structural motifs within a protein (e.g. α-helices and ß-pleated sheets)

Tertiary structure: arrangement of motifs of the secondary structure into domains (compact globular
structures)

Quaternary structure: the 3D structure of a multimeric protein, composed of several subunits
Distinguish between an α-helix and a ß-pleated sheet and appreciate the bonds that stabilise their
formation
α-helices:

Hydrogen bonds between the C=O group of 1 residue and the N–H group of another residue stabilise
the helix

Side chains of individual amino acids project out from within the helix

Right-handed helices are more common since L-enantiomers of amino acids are used in proteins

Proline: when proline is joined to a polypeptide chain, it loses its N–H group
This prevents the N atom from hydrogen bonding with the C=O group of another residue within the
helix
The helical conformation is distorted and kinked
ß-pleated sheets:

Hydrogen bonds between the C=O and N–H groups of 2 or more ß-strands stabilise the sheet

C=O and N–H groups project out perpendicular to the line of the backbone

Parallel ß-pleated sheet: alternate ß-strands run in the same direction

Anti-parallel ß-pleated sheet: alternate ß-strands run in opposite directions

Pleating allows for the best alignment of the hydrogen-bonded C=O and N–H groups
Appreciate the different types of bond that combine to stabilise a particular protein conformation
Covalent bonds: 2 atoms share electrons

The strongest bonds within a protein

Exist in the primary structure

May exist as disulphide bridges:
Disulphide bridges occur when cysteine side chains within a protein are oxidised
This results in a covalent link between the 2 amino acids
–CH2-SH + HS-CH2–  –CH2-S-S-CH2–
Hydrogen bonds: 2 atoms with partial negative charges share a partially positively charged hydrogen atom

May occur either between atoms on different Side chains and the backbone of the protein or between
water molecules
Ionic interactions: electrostatic attractions between charged side chains

Relatively strong bonds, particularly when the Side chains are within the interior of the protein and
excluded from water

The majority of charged Side chains are at the surface of a folded protein, where they can be
neutralised by counter-ions (e.g. salts)
Van der Waals forces: electrostatic attractions between 2 atoms due to the fluctuating electron cloud
surrounding each atom, which has a temporary electric dipole

Weak and transient forces

Due to the sheer number of Van der Waals interactions within a protein, they play a large part in the
overall conformation of a protein

Van der Waals radius: the appropriate distance required for Van der Waals attractions
This varies for different atoms, based on the size of the electron cloud

If the 2 electron clouds of adjacent atoms are quite close: the transient dipole in 1 atom can induce a
complementary dipole in another atom, with weak attractive properties

If the 2 electron clouds of adjacent atoms are too close: there are repulsive forces between the 2
atoms due to the electrons
Hydrophobic interactions:

Pack hydrophobic side chains into the interior of the protein

This creates a hydrophobic core and a hydrophilic surface
Give examples of the post translational modifications of amino acids, with reference to glycosylation,
hydroxylation and carboxylation
Amino acids may be modified following protein synthesis
Hydroxylation:

Proline  hydroxyproline
This requires: prolyl hydroxylase and vitamin C

Hydroxyproline is present in collagen fibres: additional hydroxyl groups help to stabilise the fibres

Scurvy: caused by vitamin C deficiency
Glycosylation:

N-linked glycosylation of asparagine: addition of sugar residues

This increases the solubility of proteins and protects them from enzymatic degradation

Primary structure motif: N-X-S/T (asparagine-any amino acid-serine/threonine)

Carbohydrate-deficient glycoprotein (CDG): associated with N-linked sugar chain transfer deficiency
Carboxylation:

Glutamate  γ-carboxyglutamate
This requires vitamin K-dependent carboxylase

Formation of γ-carboxyglutamate residues within several proteins of the blood clotting cascade is
critical for their normal function: it increases their calcium binding capability

Warfarin (anticoagulant): inhibits the above carboxylation reaction
Lecture summary:

Proteins are chains of amino acids linked by peptide bonds which have evolved to fulfil specific
functions within the cell

Such functions rely upon the conformation (3D structure) of the protein which is determined by a
variety of forces

The α-helix and ß-pleated sheet are the two staple motifs that define the conformation of a protein

The nature of the amino acid side chain dictates its position within the conformation of the protein

Post-translational modifications of proteins add more diversity to protein structure
Metabolism 2: Energetics and Enzymes
Define the 1st and 2nd Laws of thermodynamics
1st Law of Thermodynamics: energy can neither be created nor destroyed; it is simply converted from one
form to another
2nd Law of Thermodynamics: in any isolated system, such as a single cell or the universe, the degree of
disorder can only increase

Entropy: the amount of disorder in a particular system

Reactions proceed spontaneously towards products with greater entropy

Biological systems are very well ordered: energy is taken from the environment and invested into
chemical reactions within the cell which maintain order
Explain the concept of free energy and how we can use changes in free energy to predict the outcome
of a reaction
Free energy (G) [kJ/mol]: the amount of energy within a molecule that can perform useful work at a
constant temperature
∆G: the amount of disorder that results from a particular reaction
For the reaction: A + B  C + D
∆G = G (C + D) − G(A + B)

A reaction can only occur spontaneously if ∆G is negative (energetically favourable reactions)

A reaction cannot occur spontaneously if ∆G is positive (energetically unfavourable reactions)
Draw the chemical structure of ATP and explain how it acts as a carrier of free energy and is used to
couple energetically unfavourable reactions
ATP:
ATP structure:

Phosphoanhydride bonds have a large negative ∆G of hydrolysis

ATP  ADP + Pi
∆G = −31 kJ/mol
Coupled reactions:

Biosynthetic pathways are generally energetically unfavourable (e.g. peptide synthesis)

They take place because they are coupled to an energetically favourable reaction

A reaction will proceed if the sum of ∆G for the overall reaction is positive

Most energetically unfavourable biochemical reactions are coupled to the hydrolysis of high-energy
phosphoanhydride bonds (e.g. ATP hydrolysis)
Example 1: glucose + fructose  sucrose
∆G = +23 kJ/mol
This reaction is coupled to ATP hydrolysis:
Glucose + ATP  glucose-1-phosphate + ADP
Glucose-1-phosphate + fructose  sucrose
∑∆G = −31 + (+23) = −8 kJ/mol; therefore it is energetically favourable
Example 2: glucose-6-phosphate + H2O  glucose + Pi ∆G = −13.8 kJ/mol
This energetically favourable reaction will not occur at a useful rate unless it is catalysed by an enzyme
N.B. enzymes do not change the value of ∆G
Example 3: glucose + oxygen  carbon dioxide + water ∆G = −2872 kJ/mol
This energetically favourable reaction results in an increase in the entropy (disorder) of the universe
It does not occur spontaneously since energy must be supplied to overcome the activation energy barrier
Describe how enzymes act as catalysts of reactions with reference to the reaction catalysed by
lysozyme
Enzyme: a protein that acts as a catalyst to induce chemical changes in other substances itself remaining
apparently unchanged by the process
Mode of action: enzymes lower the activation energy barriers that impede chemical reactions from taking
place

1 or more substrates bind to the enzyme tightly at the active site

The enzyme arranges the substrate(s) in such a way that certain bonds are strained

Key residues within the enzyme participate in either the making or breaking of bonds by altering the
arrangement of electrons within the substrate(s)
This can either take the form of oxidation or reduction reactions
Transition state: the particular conformation of the substrate in which atoms of the molecule are
rearranged both geometrically and electronically so that the reaction can proceed

Enzymes work by bending substrates in such a way that the bonds to be broken are stressed: the
substrate molecule resembles the transition state
Lysozyme: a component of tears and nasal secretions, involved in defence against bacteria
Lysozyme catalyses the hydrolysis of sugar molecules within bacterial cell walls: this results in lysis of
bacteria

Lysozyme hydrolyses alternating polysaccharide copolymers of N-acetyl glucosamine (NAG) and Nacetyl muramic acid (NAM)

Lysozyme hydrolyses the ß-1,4 glycosidic bond which links C1 of NAM to C4 of NAG
Mechanism of action:
1.
Glu35 protonates the oxygen in the glycosidic bond: this breaks the glycosidic bond
2.
Glu35 deprotonates a water molecule: this forms a hydroxide ion
3.
Asp52 stabilises the positive charge on the transition state
4.
The hydroxide ion attacks the transition state and adds an –OH group
5.
Glu35 and Asp52 are in their original state to continue catalysis
Optimum pH for lysozyme: 5.0
At pH 5.0:

Asp52 is deprotonated (–COO−)

Glu35 is protonated (–COOH)
This is essential for lysozyme function
Describe how oxidation and reduction involve the transfer of electrons
Oxidation: loss of electrons
Reduction: gain of electrons

Since the cellular environment is generally aqueous, often when a molecule gains an electron, it
simultaneously gains a proton
Outline the reaction catalysed by glucose-6-phosphatase and explain what clinical symptoms are
linked to inherited deficiencies of this enzyme
In the liver: glycogen  glucose-6-phosphate  glucose
Glucose-6-phosphate + H2O  glucose + Pi
This reaction is catalysed by glucose-6-phosphatase

Glucose-6-phosphatase is predominantly a liver enzyme

It catalyses the above reaction when blood glucose levels are low and releases glucose from the liver
into the bloodstream
Glucose-6-phosphatase deficiency (Von Gierke’s disease)
Symptoms:

Low blood sugar levels

Slow growth

Large livers

Short stature
Outline the differences between lock and key and induced fit models of substrate-enzyme
interactions
Lock and key model: the shape of the substrate is complementary to that of the active site of the enzyme

This model explains the specificity of most enzymes for a single substrate
Induced fit model: the substrate induces a change in the conformation of the enzyme which results in the
formation of the active site; upon release of the products the enzyme reverts back to its original
conformation

This is the correct model since proteins generally possess a degree of flexibility necessary for function

Crystallographic analysis of enzymes supports this model
Describe graphically, the effects of a) substrate concentration, b) temperature and
c) pH on enzyme catalysed reactions
Effect of substrate concentration:
Enzyme kinetics:
Michaelis constant (KM): the concentration of substrate at which a given enzyme works at half its maximal
velocity (vmax)
KM is useful to compare the strength of enzyme-substrate complexes:

Low KM value: tight binding within the enzyme-substrate complex

High KM value: weak binding within the enzyme-substrate complex
At vmax the rate of product formation depends on the turnover number (i.e. how rapidly the substrate can
be processed)
Lineweaver-Burk Plot: a double reciprocal plot of 1/V against 1/[S]
Effect of temperature:
Catalysis increases as temperature is increased
Each enzyme has an optimum temperature; above this the enzyme’s conformation is denatured
Effect of pH:
The above graph is typical of most enzymes: they have an optimum pH at which the catalytic side chains
are in the correct ionisation state
Illustrate the role of the coenzyme NAD in the reactions catalysed by glyceraldehyde 3-phosphate
dehydrogenase, lactate dehydrogenase and malate dehydrogenase, referring to the biochemical
changes involved in its reduction to NADH
NAD+: a cofactor for dehydrogenation reactions

It is a coenzyme which only functions after binding to a protein

Dehydrogenases catalyse dehydrogenation reactions:
RCH(OH)R’ + NAD+  RCOR’ + NADH + H+

NAD+ catalyses dehydrogenation of substrates by accepting a hydrogen atom and 2 electrons:
NAD+ + H+ + 2e-  NADH
Glyceraldehyde 3-phosphate dehydrogenase catalyses…
Glyceraldehyde 3-phosphate + NAD+ + Pi  1,3-bisphosphoglycerate + NADH + H+

The substrate is oxidatively phosphorylated
Lactate dehydrogenase catalyses…
Pyruvate + NADH + H+  lactate + NAD+

Pyruvate is converted into lactate by anaerobic respiration during intense exercise

This generates free NAD+ which is needed by muscle for other reactions

Lactate is transported from muscles to the liver by the bloodstream

The liver has high NAD+ levels which can be used by lactate dehydrogenase to regenerate pyruvate
Malate dehydrogenase catalyses…
Malate + NAD+  oxaloacetate + NADH + H+
Lecture summary:

Energetically unfavourable reactions can occur by coupling them to energetically favourable
reactions, generally involving the hydrolysis of high-energy phosphoanhydride bonds

Enzymes are specific biological catalysts which increase the rate of biochemical reactions by lowering
the activation energy barriers that impede chemical reactions

Enzymes are sensitive to extremes of temperature and pH, as they are proteins
Metabolism 3: Metabolic Pathways and ATP Production I
Sketch a cartoon of the three stages of cellular metabolism that convert food to waste products in
higher organisms, illustrating the cellular location of each stage
Cellular metabolism: energy is liberated from food molecules to provide energy
3 food molecules are used by cells:
1.
Polysaccharides  simple sugars
2.
Proteins  amino acids
3.
Fats  fatty acids and glycerol
3 stages of metabolism:
1.
Digestion: enzymes liberate small molecules in the small intestines
2.
Cellular metabolism I: small molecules are oxidised to generate ATP and NADH in the cell cytosol
3.
Cellular metabolism II: small molecules are oxidised to generate ATP within the mitochondria
Glucose combustion: single-step reaction

The relatively large activation energy is overcome by a heat source

Free energy is released as heat
Glucose metabolism: multi-step reaction

The relatively small activation energies of each step are overcome by enzymes and body temperature

Free energy liberated is invested into carrier molecules, such as ATP

This reaction is ~50% efficient
Outline the metabolism of glucose by the process of glycolysis, listing the key reactions, in particular
those reactions that consume ATP and those that generate ATP
Glycolysis: an anaerobic process which occurs in the cell cytoplasm
1.
Glucose + ATP  glucose 6-phosphate + ADP + H+
Hexokinase
Glucose is committed to further reactions and it is trapped inside the cell (due to negative charge)
2.
Glucose 6-phosphate  fructose 6-phosphate
Phosphoglucose isomerase
3.
Fructose 6-phosphate + ATP  fructose 1,6-bisphosphate + ADP
Phosphofructokinase
Fructose 1,6-bisphosphate is a highly symmetrical, high energy compound
4.
Fructose 1,6-bisphosphate  glyceraldehyde 3-phosphate + dihydroxyacetone phosphate
Aldolase
5.
Dihydroxyacetone phosphate  glyceraldehyde 3-phosphate
Triose phosphate isomerase
6.
Glyceraldehyde 3-phosphate + NAD+ + Pi  1,3-bisphosphoglycerate + NADH
Glyceraldehyde 3-phophate dehydrogenase
7.
1,3-bisphosphoglycerate + ADP  3-phosphoglycerate + ATP
Phosphoglycerate kinase
8.
3-phosphoglycerate  2-phosphoglycerate
Phosphoglycerate mutase
9.
2-phosphoglycerate  phosphoenolpyruvate + H2O
Enolase
10. Phosphoenolpyruvate + ADP  pyruvate + ATP
Pyruvate kinase
Net output of glycolysis per glucose molecule: 2 pyruvate; 2 ATP; 2 NADH + H+
Substrate-level phosphorylation: ATP is produced by the direct transfer of a high-energy phosphate group
from an intermediate substrate in a biochemical pathway to ADP, such as occurs in glycolysis
Oxidative phosphorylation: ATP is produced using energy derived from the transfer of electrons in an
electron transport system
Distinguish between the aerobic and anaerobic metabolism of glucose with reference to the enzymes
involved and the comparative efficiencies of each pathway with respect to ATP generation
Anaerobic metabolism of glucose:
Alcoholic fermentation: characteristic of yeasts
1.
Pyruvate + H+  acetaldehyde + CO2
Pyruvate decarboxylase
2.
Acetaldehyde + NADH + H+  ethanol + NAD+
Alcohol dehydrogenase
Lactate production: characteristic of mammalian muscle during intense activity when oxygen is a limiting
factor
Pyruvate + NADH + H+  lactate + NAD+
Lactate dehydrogenase
Anaerobic metabolism of glucose involves a dehydrogenase: this allows NAD+ to be regenerated to allow
glycolysis to continue in conditions of oxygen deprivation

NAD+: required for the dehydrogenation of glyceraldehyde 3-phosphate

Oxygen deprivation: conditions in which the rate of NADH formation by glycolysis is greater than the
rate of oxidation of NADH by the respiratory chain
Aerobic metabolism of glucose:
Acetyl CoA production:
Pyruvate + HS-CoA + NAD+  acetyl CoA + CO2 + NADH
Pyruvate dehydrogenase complex

This series of reactions occur in the mitochondrial matrix

This series of reactions proceeds through the pyruvate dehydrogenase complex

Acetyl CoA is committed to enter the TCA cycle
Aerobic metabolism of glucose yields considerably more ATP than anaerobic metabolism
Anaerobic metabolism yields 2 ATPs; whereas complete aerobic metabolism can yield 38 ATPs
Describe the reactions catalysed by lactate dehydrogenase and creatine kinase and explain the
diagnostic relevance of their appearance in plasma
Lactate dehydrogenase (LDH): catalyses the interconversion of pyruvate and lactate
LDH is present in the heart, liver, kidneys, skeletal muscle, brain and lungs
Elevated LDH levels can be used to diagnose several disorders including:

Stroke

Myocardial infarction

Liver disease (e.g. hepatitis)

Muscle injury

Muscular dystrophy

Pulmonary infarction
Creatine kinase:
Creatine kinase catalyses the following reaction:
Creatine phosphate + ADP + H+  creatine + ATP
∆G = −43.1 kJ/mol (c.f. −31 kJ/mol for ATP)
A large reservoir of creatine phosphate is present in muscle: it buffers the demand for phosphate (which is
required for ATP synthesis)
When a muscle is damaged, creatine kinase leaks into the bloodstream
Either total creatine kinase levels (70% accurate) or the level of the tissue specific isoform (90% accurate)
can be measured to determine which tissue has been damaged
Elevated creatine kinase levels can be used to:

Diagnose myocardial infarction

Determine the extent of muscular disease

Evaluate the cause of chest pain

Help to identify carriers of Duchene muscular dystrophy
Outline the oxidative decarboxylation reaction catalysed by pyruvate dehydrogenase, with reference
to the product and the five co-enzymes required by this enzyme complex
Pyruvate forms acetyl CoA via the pyruvate dehydrogenase complex
The pyruvate dehydrogenase complex is gigantic: it consists of 3 enzymes and 5 cofactors (60
polypeptides)
Enzyme
Prosthetic group
Reaction catalysed
E1
Pyruvate decarboxylase
TPP
Oxidative decarboxylation of pyruvate
E2
Lipoamide reductase-transacetylase
Lipoamide
Transfer of acetyl group to CoA
E3
Dihydrolipoyl dehydrogenase
FAD
Regeneration of oxidised lipoamide
Cofactors: thiamine pyrophosphate (TPP), lipoamide, FAD, CoA and NAD+

Prosthetic group: a cofactor which forms a permanent part of the complex with an enzyme

Coenzyme: a cofactor which binds reversibly to its corresponding enzyme
o
CoA: a coenzyme for E2
o
NAD+: a coenzyme for E3
Thiamine pyrophosphate (TPP):

Derivative of thiamine (vitamin B1)

It readily loses a proton: this forms a carbanion which attacks that of pyruvate to form hydroxyethylTPP

Thiamine deficiency causes Beriberi
Symptoms include: damage to the peripheral nervous system, muscle weakness and reduced cardiac
output
Lipoamide:

The dithiol group undergoes redox reactions

The long flexible arm allows the dithiol group to swing between active sites within the complex

Arsenite (AsO33−) and mercury inhibit the pyruvate dehydrogenase complex since they have a high
affinity for sulphydryl groups which are found in reduced lipoamide
Flavine adenine dinucleotide (FAD):

FAD + 2e- + 2H+  FADH2
Mechanism of the pyruvate dehydrogenase complex:
1.
E1: decarboxylation of pyruvate
Pyruvate + carbanion of TPP  hydroxyethyl TPP + CO2
2.
E1: oxidation of hydroxyethyl TPP
Hydroxyethyl TPP + lipoamide  acetyllipoamide + carbanion of TPP
3.
E2: transfer of acetyl group from acetyllipoamide to CoA
Acetyllipoamide + CoA  acetyl CoA + dihydrolipoamide
4.
E3: regeneration of oxidised lipoamide (dehydrogenation)
Dihydrolipoamide + FAD  lipoamide + FADH2
5.
Regeneration of FAD:
FADH2 + NAD+  FAD + NADH + H+
Overall reaction: pyruvate + HS-CoA + NAD+  acetyl CoA + CO2 + NADH
Describe the different processes by which the fatty acid palmitate and the amino acid alanine are
converted into acetyl-CoA
Fatty acid metabolism: occurs in the mitochondria
Acetyl CoA is produced from both polysaccharides and fats within the mitochondria:
1.
Polysaccharides  sugars  glucose  pyruvate  acetyl CoA
2.
Fats  fatty acids  acetyl CoA
Fatty acids are the most compact fuel since they are fully reduced (i.e. the carbon skeleton is saturated
with hydrogen)
Fatty acid metabolism yields several times the useful energy compared with carbohydrates
Mechanism of fatty acid metabolism:
1.
The fatty acid is converted into an acyl CoA species:
Fatty acid + ATP + HS-CoA  acyl CoA + AMP + PPi
Acyl CoA synthetase
2.
The acyl CoA species undergoes ß-oxidation:
Cn acyl CoA + NAD+ + FAD + H2O + CoA  Cn−2 acyl CoA + acetyl CoA + NADH + FADH2
E.g. palmitoyl CoA + NAD+ + FAD + H2O + CoA  myristyl CoA + acetyl CoA + NADH + FADH2
ß-oxidation: involves a sequence of dehydration, hydration, oxidation and thiolysis reactions

Each ß-oxidation reaction removes 2 carbons from the acyl CoA and produces 1 acetyl CoA
Metabolism of palmitic acid [C16]: involves 7 ß-oxidation reactions
1. Palmitic acid + ATP + HS-CoA  palmitoyl CoA + AMP + PPi
2. Palmitoyl CoA + 7 NAD+ + 7 FAD + 7 H2O + 7 CoA  8 acetyl CoA + 7 NADH + 7 FADH2
Amino acid metabolism: occurs in the liver
The pathway of amino acid metabolism depends upon the number of carbon atoms in the amino acid

C3 amino acids  pyruvate

C4 amino acids  oxaloacetate

C5 amino acids  α-ketoglutarate
Transamination: alanine [C3] undergoes transamination
Alanine + α-ketoglutarate  pyruvate + glutamate
Alanine aminotransferase

Pyruvate can enter the TCA cycle

Glutamate is converted to α-ketoglutarate by glutamate dehydrogenase
This generates NH4+ ions which are ultimately converted to urea

Persistently elevated levels of alanine aminotransferase are indicative of hepatic disorders (e.g.
Hepatitis C)
Metabolism 4: Metabolic Pathways and ATP Production II
Outline the Krebs or TCA (tricarboxylic acid cycle) with particular reference to the steps involved in the
oxidation of acetyl Co-A and the formation of NADH and FADH2 and the cellular location of these
reactions
Acetyl CoA: possesses a high-energy thioester bond (C-S) between the acetyl group and CoA

The thioester bond is readily hydrolysed; therefore acetyl CoA can donate its acetyl group to other
molecules (i.e. oxaloacetate in the Krebs cycle)
Krebs cycle: an aerobic process which occurs in the mitochondrial matrix
1.
Oxaloacetate + acetyl-CoA  citrate + HS-CoA + H+
Citrate synthase
2.
Citrate  isocitrate
Aconitase
3.
Isocitrate + NAD+  α-ketoglutarate + NADH + H+ + CO2
Isocitrate dehydrogenase
4.
α-ketoglutarate + NAD+ + HS-CoA  succinyl CoA + NADH + H+ + CO2
α-ketoglutarate dehydrogenase complex
5.
Succinyl CoA + GDP + Pi + H2O  succinate + GTP* + HS-CoA
Succinyl CoA synthetase
6.
Succinate + FAD  fumarate + FADH2
Succinate dehydrogenase
7.
Fumarate + H2O  malate
Fumerase
8.
Malate + NAD+  oxaloacetate + NADH + H+
Malate dehydrogenase
Net output per turn of the cycle: 1 GTP, 3 NADH, 1 FADH2 and 2 CO2
*GTP (guanosine triphosphate):

GTP can act as a phosphoryl donor in protein synthesis or in signal transduction processes (e.g. G
protein-coupled receptors)

The γ-phosphate group of GTP can be transferred to ADP to generate ATP:
GTP + ADP  GDP + ATP
Nucleoside diphosphokinase
Cellular location of the Krebs cycle:

Krebs cycle enzymes: soluble proteins which are located in the mitochondrial matrix, except for…

Succinate dehydrogenase: an integral membrane protein which is embedded in the inner
mitochondrial membrane
N.B. The Krebs cycle only occurs under aerobic conditions since reduced cofactors are required: these must
be reoxidised via oxidative phosphorylation
Reoxidation of reduced cofactors via oxidative phosphorylation generates ATP:

Re-oxidation of NADH  3 ATP

Re-oxidation of FADH2  2 ATP
Therefore overall the Krebs cycle generates: 3 NADH + 1 FADH2 + 1 GTP = 12 ATP per acetyl CoA
Calculate the theoretical maximum yield of ATP per glucose molecule oxidized by aerobic respiration
and compare this to the theoretical maximum yield of ATP per molecule of palmitic acid
Theoretical maximum yield of ATP…
Per molecule of glucose oxidised by aerobic respiration:
Glycolysis:
Glucose  2 pyruvate + 2 ATP + 2 NADH

Direct production of 2 ATP

Oxidative phosphorylation: 2 NADH  6 ATP
Total: 8 ATP
Pyruvate dehydrogenase complex:
2 pyruvate + 2 NAD+ + CoA  2 acetyl CoA + 2 NADH + 2 CO2

Oxidative phosphorylation: 2 NADH  6 ATP
Total: 6 ATP
Krebs cycle:
2 acetyl CoA  6 NADH + 6 H+ + 2 FADH2 + 2 GTP

Direct production of 2 GTP ≡ 2 ATP

Oxidative phosphorylation: 6 NADH  18 ATP

Oxidative phosphorylation: 2 FADH2  4 ATP
Total: 24 ATP
Grand total: 8 + 6 + 24 = 38 ATP
Per molecule of palmitic acid:
1st step of ß-oxidation: conversion of fatty acid  acyl CoA species
Palmitic acid + ATP + HS-CoA  palmitoyl CoA + AMP + PPi
Total: − 2 ATP (2 high-energy phosphoanhydride bonds are used and this is the equivalent of 2 ATPs)
Subsequent steps of ß-oxidation:
Palmitoyl CoA + 7 NAD+ + 7 FAD + 7 H2O + 7 CoA  8 acetyl CoA + 7 NADH + 7 FADH2

7 NADH  21 ATP

7 FADH2  14 ATP

8 acetyl CoA  96 ATP
Total: 131 ATP
Grand total: 21 + 14 + 96 − 2 = 129 ATP
Outline the glycerol phosphate shuttle and the malate-aspartate shuttle, in particular stating why
these mechanisms are required
NAD+ needs to be regenerated by oxidative phosphorylation in order for glycolysis to continue since NAD+
is required for dehydrogenation reactions
NADH cannot diffuse directly from the cytosol into the mitochondrial matrix since the inner mitochondrial
membrane is impermeable to NADH and NAD+
There are 2 mechanisms by which high-energy electrons can be transferred from cytosolic NADH across
the mitochondrial membrane
1) Glycerol phosphate shuttle: found in skeletal muscle and in the brain
In the cytosol:
Dihydroxyacetone phosphate + NADH  glycerol 3-phosphate + NAD+
Cytosolic glycerol 3-phosphate dehydrogenase (GPD)

Cytosolic GPD transfers electrons from NADH to form glycerol 3-phosphate

Glycerol 3-phosphate diffuses from the cytosol into the mitochondria
In the mitochondria:
Glycerol 3-phosphate + FAD  dihydroxyacetone phosphate + FADH2
Mitochondrial glycerol 3-phosphate dehydrogenase (membrane-bound GPD)

Mitochondrial GPD transfers electrons from glycerol 3-phosphate to FAD
2) Malate-aspartate shuttle: found in the heart, liver and kidneys
The malate-aspartate shuttle is mediated by 2 membrane carriers and 4 enzymes
Net reaction: cytosolic NADH + mitochondrial NAD+  cytosolic NAD+ + mitochondrial NADH
In the cytosol:
Oxaloacetate + NADH  malate + NAD+
Cytosolic malate dehydrogenase (MDH)

Cytosolic MDH transfers electrons from NADH to oxaloacetate to form malate

Malate is transported into the mitochondrial matrix by an α-ketoglutarate transporter
In the mitochondria:
Malate + NAD+  oxaloacetate + NADH
Mitochondrial malate dehydrogenase (MDH)

Mitochondrial MDH catalyses reoxidation of malate by NAD+ to form oxaloacetate
Understand the concept of transamination with reference to the malate-aspartate shuttle
Oxaloacetate cannot readily cross the inner mitochondrial membrane:

It accumulates in the mitochondrial matrix

It is depleted from the cytosol
A transamination reaction is necessary to convert oxaloacetate to aspartate which can be transported
across the mitochondrial membrane by a glutamate/aspartate transporter
Transamination: a reaction in which an amino group is transferred from an amino acid to a keto acid,
thereby forming a new pair of amino and keto acids

One of the pairs is almost always glutamate (an amino acid) and α-ketoglutarate (a keto acid)
In the mitochondria:
Oxaloacetate + glutamate  aspartate + α-ketoglutarate
Mitochondrial aspartate aminotransferase

Transamination: glutamate transfers an amino group to oxaloacetate to form aspartate

Aspartate is transported into the cytosol by a glutamate/aspartate transporter
In the cytosol: the same transamination reaction occurs but in reverse
Aspartate + α-ketoglutarate  oxaloacetate + glutamate
Cytosolic aspartate aminotransferase
Explain in general terms the relationship between TCA intermediates and those pathways involved in
amino acids synthesis and breakdown
Amino acid breakdown: the amino group is removed and the carbon skeleton is used to produce Krebs
cycle intermediates or glucose

Degradation of all 20 amino acids produces 7 Krebs cycle intermediates: pyruvate, acetyl CoA,
acetoacetyl CoA, α-ketoglutarate, succinyl CoA, fumarate and oxaloacetate
Give two examples of the use of NADPH in reductive biosynthesis
Biosynthesis: the synthesis of macromolecules from smaller, simple molecules

Glycolysis and the Krebs cycle provide intermediates for many biosynthetic reactions

Krebs cycle intermediates which are drawn off for biosynthesis must be replenished
E.g. if oxaloacetate is removed, acetyl CoA cannot enter the Krebs cycle; therefore glycolysis stops

Anaplerotic reactions: enzyme-catalysed reactions which regenerate Krebs cycle intermediates
Pyruvate + CO2 + ATP + H2O  oxaloacetate + ADP + Pi + 2 H+
Pyruvate carboxylase
NADP+: nicotinamide adenine dinucleotide phosphate

Similar structure to NAD+: it differs by a single phosphate group attached to one of the ribose rings

The additional phosphate group gives NADP+ a different conformation to NAD+; therefore NADP+
binds to different enzymes than NAD+

NADP+ can accept 2 electrons and 1 proton (i.e. a hydride ion) like NAD+:
NADP+ + H+ + 2e−  NADPH

The hydride ion is held by a high-energy linkage; therefore it can be easily transferred

NADPH participates in anabolic reactions; whereas NADH participates in catabolic reactions
This allows electron transport in catabolism to be kept separate from that of anabolism

NADPH is a cofactor for reduction in:
o
Biosynthesis of RNA
o
Biosynthesis of cholesterol: NADPH catalyses the final reaction of cholesterol synthesis
The C=C bond in 7-dehydrocholesterol is reduced by the transfer of a hydride ion from NADPH
Metabolism 5: Mitochondria and Oxidative Phosphorylation
Outline the proposed evolutionary origins of mitochondria
Endosymbiotic theory: mitochondria are believed to be the evolutionary descendents of a prokaryote
which formed a symbiotic relationship with the ancestors of eukaryotic cells
Following this many genes needed for mitochondrial function were translocated to the eukaryotic genome
Evidence for the theory:

Mitochondria can only arise from pre-existing mitochondria

Mitochondria have their own genome: a single circular DNA molecule with no associated histones
This resembles the genome of prokaryotes
The mitochondrial genome is similar to that of Rickettsia prowazekii

Mitochondria have their own protein-synthesising machinery: 70S ribosomes
This resembles the protein-synthesising machinery of prokaryotes

The first amino acid of the transcript is fMet (as with prokaryotes), not Met (as with eukaryotes)

Antibiotics (e.g. streptomycin) block protein synthesis in both prokaryotes and mitochondria, and do
not interfere with protein synthesis in the cytoplasm of eukaryotes
Draw a cross sectional representation of a mitochondrion, and label its component parts
Mitochondrial structure: a rod-shaped organelle with 2 membranes

Outer membrane: limits the size of the mitochondrion

Intermembrane space: the space between the inner and outer mitochondrial membranes

Inner membrane: has many cristae (infoldings that project into the mitochondrial matrix)
Cristae: invaginations of the inner membrane which increase the surface area upon which oxidative
phosphorylation can occur and enhance ATP production

Matrix: the space enclosed by the inner membrane; it contains the Krebs cycle enzymes
In some cells mitochondria are mainly distributed where ATP is rapidly consumed
E.g. between the myofibrils of muscle cells; wrapped around the flagellum of sperm
Outline the chemiosmotic theory
Oxidative phosphorylation: ATP synthesis is driven by the transfer of high-energy electrons from reduced
coenzyme to molecular O2 via the ETC

Within mitochondria reduced coenzyme is reoxidised by molecular oxygen:
NADH + H+ + ½ O2  NAD+ + H2O
∆G = −220 kJ/mol
FADH2 + ½ O2  FAD + H2O
∆G = −167 kJ/mol

Energy released by the reoxidation of reduced coenzyme can be used to generate several
phosphoanhydride bonds (∆G ATP hydrolysis = −31 kJ/mol)

Some of the energy released by the reoxidation of reduced coenzyme is recovered by components of
the ETC and used to synthesise ATP
Chemiosmotic hypothesis of oxidative phosphorylation:
1.
Protons are translocated from the mitochondrial matrix into the intermembrane space
The ETC provides the energy for active transport of protons
2.
Protons diffuse back into the mitochondrial matrix through a specific proton channel which is coupled
to ATP synthase: flow of protons back into the matrix is coupled to ATP synthesis
The proton motive force which drives protons back into the mitochondrial matrix consists of a pH gradient
and a transmembrane electric potential
Describe the electron transport chain in mitochondria with reference to the functions of coenzyme Q
(ubiquinone) and cytochrome c
Electron transport chain (ETC): a chain of 3 membrane complexes and 2 mobile carriers
Membrane complexes: integral membrane proteins
1.
NADH dehydrogenase complex
2.
Cytochrome b-c1 complex
3.
Cytochrome oxidase complex
Mobile carriers: fixed in the membrane
1.
Ubiquinone (coenzyme Q)
2.
Cytochrome c
Ubiquinone: a mobile electron carrier

Ubiquinone can accept either 1 or 2 electrons, together with protons (from solution)

It passes electrons to cytochrome b-c1 complex

It has a hydrophobic tail: this confines it to the lipid bilayer of the membrane
Succinate dehydrogenase: an integral membrane protein

Succinate dehydrogenase is embedded in the inner surface of the inner mitochondrial membrane: this
allows it to pass its electrons directly to ubiquinone via FAD

Ubiquinone is the entry point for electrons donated by FADH2
Therefore fewer protons are pumped into the intermembrane space compared with NADH and less
ATP is produced
Cytochrome oxidase: involved in the final step of electron transfer

Cytochrome oxidase receives 4 electrons and 4 protons from cytochrome c
4e− + 4H+ + O2  2H2O

It pumps 4 additional protons into the intermembrane space: this enhances the proton gradient

Molecular O2 is an ideal terminal electron acceptor since it has a high affinity for electrons: this
provides a driving force for oxidative phosphorylation

Proteins of the ETC accept electrons and protons from the aqueous solution

Each unit of the ETC has a higher affinity for electrons than the previous unit

Electron transfer along the ETC is energetically favourable: as electrons progress along the chain, they
lose energy

As electrons pass through each membrane complex, protons are pumped into the intermembrane
space
Redox reactions:

Redox reaction: an electron transfer reaction involving a reduced substrate and an oxidised substrate

Redox couple: a substrate that can exist in both oxidised and reduced forms
E.g. NAD+/NADH + H+ and FAD/FADH2

Redox potential: the ability of a redox couple to either accept or donate electrons

Standard redox potential (E’o): determined experimentally by allowing a reaction to reach equilibrium
and measuring the resultant e.m.f.

E’o conditions for biological systems: 25°C; pH 7.0; reactant concentrations of 1.0M

Standard hydrogen electrode: used as a reference electrode
Describe how ATP synthase is able to generate and utilise ATP respectively, with reference to its
structure
ATP synthase: a multimeric enzyme complex that catalyses the synthesis of ATP from ADP and Pi
Structure of ATP synthase:

2 parts: F0 and F1
F0 unit: membrane-bound
F1 unit: projects into the matrix

Structure of the F0 unit: a, b and c subunits

Structure of the F1 unit: α, ß and γ subunits

The c subunits are arranged into a disc which is fixed to the γ subunit

The α and ß subunits cannot rotate since they are locked in a fixed position by the b subunit

The b subunit is anchored to the a subunit in the membrane
Mechanism of ATP synthesis:
1.
Protons flow across the inner mitochondrial membrane via a pore
2.
This causes the disc of c subunits to rotate
3.
This in turn causes the attached γ subunit to rotate
4.
The catalytic portions of the ß subunits undergo conformational changes: this changes their affinities
for ADP and ATP
5.
Torsional energy flows from the catalytic portion to the bound ADP and Pi: this forms ATP
Binding change mechanism: the active site of a ß subunit cycles between 3 states

Open state: ADP and Pi bind to the active site

Loose binding state: the enzyme closes up around the molecules and binds them loosely

Tight binding state: the enzyme changes shape and forces the molecules together to form ATP
ATP synthesis is a reversible reaction: ADP + Pi  ATP
Position of equilibrium depends on the direction of proton flow through ATP synthase:

Protons flowing into the matrix  ATP synthesis

Protons flowing into the intermembrane space  ATP hydrolysis
Explain why carbon monoxide, cyanide, malonate and oligomycin are poisonous in terms of their
effects on specific components of the electron transport chain
If oxidative phosphorylation is disrupted, cells become rapidly depleted of ATP and may die

Lack of oxygen causes failure of oxidative phosphorylation: hypoxia (diminished oxygen) and anoxia
(complete lack of oxygen)
Drug/Toxin
Effect on the ETC
Cyanide (CN−)
Bind to the ferric (Fe3+) form of the haem group in cytochrome oxidase with high affinity
Azide (N3−)
Cyanide is supertoxic: i.e. ingestion of a few drops can be lethal
Carbon monoxide
Binds to the ferrous (Fe2+) form of the haem group in cytochrome oxidase with high affinity
Malonate
Acts as a competitive inhibitor of succinate dehydrogenase: it competes with succinate
Oligomycin
Inhibits oxidative phosphorylation: it binds to the stalk of ATP synthase and blocking the flow
of protons through it
Dinitrophenol
Causes metabolic uncoupling: it transports protons across the mitochondrial membrane
Describe how oxidative phosphorylation can be measured experimentally
Oxygen electrode: measures the oxygen concentration of a solution

2 electrodes: platinum cathode and silver anode

The reaction chamber is isolated from the electrode compartment by a Teflon membrane

Teflon membrane: permeable to oxygen

A small voltage is applied between the electrodes (0.6 V)

Oxygen is reduced at the platinum cathode:
O2 + 4H+ + 4e−  2H2O

The circuit is completed at the silver anode, which is slowly corroded by the KCl electrolyte:
Ag + Cl−  AgCl + e−

The resulting current is proportional to the oxygen concentration in the reaction chamber
The oxygen electrode can be used to measure oxidative phosphorylation:

Incubate a suspension of mitochondria from homogenised tissue with an isotonic medium containing
substrate (e.g. succinate and Pi)

Add ADP: this causes a sudden burst of oxygen uptake since ADP is converted into ATP
Understand the difference between oxidative phosphorylation and substrate level phosphorylation
Substrate level phosphorylation: the synthesis of ATP by transfer of a high-energy Pi from an intermediate
substrate in a biochemical pathway to ADP
Oxidative phosphorylation: the synthesis of ATP from ADP and Pi using energy from the transfer of highenergy electrons from reduced coenzyme to O2 via the ETC
Metabolism 6: Lipids and Membranes
Describe the structure of: fatty acids, triglycerides, phospholipids, cholesterol and sphingomyelin
Fatty acids: the simplest lipids

Structure: hydrocarbon chain + carboxyl group

Fatty acids are amphipathic: i.e. they possess both hydrophobic and hydrophilic regions
Hydrophilic head: the carboxyl group (undergoes esterification reactions)
Hydrophobic tail: the hydrocarbon chain (may be either saturated or unsaturated)

Function: fatty acids are constituents of more complex lipids (phospholipids and sphingolipids)
Triacylglycerols (triglycerides):

Structure: 3 fatty acid chains + glycerol linked by ester bonds

Ester bonds: the carboxyl group on each fatty acid is esterified to a hydroxyl group on glycerol

Function: triacylglycerols are metabolic energy stores
Membrane structure:
2 membrane types:
1.
Plasma membranes: define the outer limit of the cell
2.
Intracellular (organelle) membranes: separate and limit the size of organelles
Fluid mosaic model of the membrane:

Membranes are 2D solutions of oriented proteins and lipids which are held non-covalently

The membrane is a continuous bilayer ~5 nm thick

Proteins are either (a) integral or (b) peripheral
o
Integral proteins: span the entire bilayer
o
Peripheral proteins: present on either side of the membrane
Lipid rafts:

Membranes are not homogeneous and are dynamic

Lipid rafts are specialised domains of the plasma membrane which are enriched in certain lipids
(glycolipids and cholesterol) and proteins
Structure of the lipid bilayer:
The lipid bilayer is composed of phospholipids, sphingolipids, glycolipids and steroids
Phospholipids:

Structure: glycerol backbone + tail (2 fatty acid chains) + head group (contains phosphate)
Fatty acid chain variables: (a) length and (b) saturation

Common phospholipids include:
Phosphatidylcholine (PC): choline head group
Phosphatidylserine (PS): serine head group
Phosphatidylethanolamine (PE): ethanolamine head group
Phospholipids in aqueous solution shield the hydrophobic core by:

Bilayer formation: in the case of amphipathic lipids (e.g. phospholipids and glycolipids)

Micelle formation: in the case of non-amphipathic lipids (e.g. triglycerides)
Sphingolipids and glycosphingolipids:

Derived from sphingosine (an amino alcohol with a long, unsaturated hydrocarbon chain)

Sphingolipid structure: a long hydrocarbon chain is linked to the amino group on sphingosine by an
amide bond

Sphingosine + fatty acid  ceramide

Ceramide + sugar  glycosphingolipid

Glycosphingolipids include cerebrosides and gangliosides
Function: structural role; recognition of molecules

Sphingomyelin: phosphocholine + ceramide
Sphingomyelin is a common membrane lipid
Give examples of how the lipid composition can differ for different cellular membranes, and indicate
the significance of this
Membrane flexibility: the ability of the membrane to bend
Membrane fluidity: the ability of lipid molecules to move in the plane of the bilayer (the monolayer)
Individual lipid molecules very rarely move from one monolayer to the other (flip-flop)
Factors affecting fluidity:



Length of the hydrocarbon tail:
o
Longer carbon chains have a greater surface area over which dispersion forces can act
o
Therefore long hydrocarbon chains have a greater tendency to aggregate  lower fluidity
Degree of saturation of the hydrocarbon tail:
o
Unsaturated fatty acids have kinked chains and the dispersion forces between the hydrocarbon
chains are less stable
o
Therefore hydrocarbon chains with a greater degree of unsaturation have a lower tendency to
aggregate  greater fluidity
Cholesterol: decreases membrane fluidity (i.e. increases membrane rigidity)
Asymmetry of membrane lipids:
Within cell membranes: lipid composition of cell membranes varies according to the cell and they type of
cell membrane (i.e. plasma membrane or intracellular membrane)
E.g. the plasma membrane of human erythrocytes has a much higher percentage of glycolipids,
phosphatidylcholine and cholesterol compared to the mitochondrial membrane of beef heart
Within bilayers:

Phospholipids and glycolipids are unevenly distributed in the extracellular and cytosolic monolayers
o
Extracellular monolayer: composed of glycolipids, sphingomyelin and phosphatidylcholine
o
Cytosolic monolayer: composed of phosphatidylserine, phosphatidylethanolamine and
phosphatidylinositol

Certain lipids are organised into microdomains within monolayers of the plasma membrane

Lipid rafts: microdomains which are enriched in sphingomyelin, glycosphingolipid, phospholipid,
cholesterol, cell-surface receptor proteins and signalling proteins
Lipid rafts act as signalling sites for signalling across the plasma membrane
Outline the pathway for synthesis of fatty acids
Fat digestion, absorption and storage:
1.
Bile salts emulsify fats
2.
Hydrolytic enzymes (secreted by the pancreas) hydrolyse fats in the small intestine
3.
Triacylglycerols are transported from the small intestine to the liver in the bloodstream via
chylomicrons
4.
Triacylglycerols are transported from the liver to muscles and the heart in the bloodstream via VLDLs
5.
Triacylglycerols are stored in adipose tissue
Fatty acid synthesis (aka lipogenesis):
Acetyl CoA: the key intermediate between fat and carbohydrate metabolism

Acetyl CoA + oxaloacetate  citrate

Acetyl CoA  fatty acids

If there is too much acetyl CoA: it is stored as fatty acids

If there is too little acetyl CoA: sources of acetyl CoA are mobilised
o
Carbohydrates are short-term stores of acetyl CoA
o
Triacylglycerols are long-term stores of acetyl CoA
Fatty acid biosynthesis: acetyl CoA  palmitate [C16]
1.
Production of malonyl CoA: acetyl CoA [C2]  malonyl CoA [C3]
Enzyme: acetyl CoA carboxylase
Cofactor: biotin
2.
Activation of acetyl CoA/malonyl CoA by acyl carrier protein (ACP): acyl CoA  acyl ACP
Cofactor: phosphopantetheine
3.
Elongation of acetyl ACP/malonyl ACP by successive addition of 2 carbons
Enzyme: fatty acid synthase (FA synthase)
This stage is very similar to fatty acid degradation by ß-oxidation: each cycle adds 2 carbons
2 enzymes are involved in fatty acid synthesis:
1.
Acetyl CoA carboxylase: this catalyses the first step
2.
FA synthase: this contains multiple activities which catalyse the elongation reactions
Overall reaction:
Acetyl CoA + 7 malonyl CoA + 14 NADPH + 14 H+  palmitate + 7 CO2 + 6 H2O + 8 HS-CoA + 14 NADP+
Further metabolism of palmitate:

Esterification: this forms triacylglycerols

Formation of other fatty acids by:
o
Desaturation: this forms unsaturated fatty acids, including monounsaturated fatty acids
o
Elongation: this forms longer chain fatty acids
Regulation of fatty acid synthesis:

Feedback inhibition of palmitoyl CoA to:
o
Acetyl CoA carboxylase
o
FA synthase
o
Pentose phosphate pathway: this is a source of NADPH
Glucose 6-phosphate  pentose phosphate + NADPH

Hormonal regulation of acetyl CoA carboxylase

Transcriptional regulation of acetyl CoA carboxylase and FA synthase:
o
Activated by insulin (and citrate)
o
Inhibited by glucagon (and adrenaline and palmitoyl CoA)
Distinguish between the pathways for synthesis and metabolism of fatty acids in terms of: substrates
and products, coenzymes used, carrier molecules, cellular location
Cellular location
Substrates
Products
Fatty acid synthesis
Fatty acid degradation
Cytosol
Mitochondrial matrix
Acetyl CoA + malonyl CoA + NADPH + H+
Fatty acid + CO2 + H2O + HS-CoA + NADP+
1.
Fatty acid + ATP + HS-CoA
2.
Acyl CoA + NAD+ + FAD + H2O + CoA
1.
Acyl CoA + AMP + PPi
2.
Acetyl CoA + NADH + FADH2
Carrier molecules
ACP
HS-CoA
Redox cofactor
NADPH (reducing agent)
NAD+ and FAD (oxidising agents)
Metabolism 7: Cholesterol
Explain the physiological functions of cholesterol in membrane stability
Structure of cholesterol:

Derivative of a saturated tetracyclic hydrocarbon

Planar, rigid structure: all cyclohexane rings are in the chair conformation

The storage form of cholesterol is acylated at position 3 (i.e. where the–OH group is usually present)
Physiological functions of cholesterol:

Regulates membrane fluidity: high cholesterol leads to reduced membrane fluidity
o
It reduces phase transitions of lipids
o
It reduces the lateral mobility of polar lipids

Precursor of steroid hormones: cortisol (a glucocorticoid), aldosterone (a mineralocorticoid),
progesterone (a progestin), oestradiol (an oestrogen) and testosterone (an androgen)

Precursor of bile salts

Signalling molecule: cholesterol is involved in lipid raft formation at the plasma membrane
Outline the synthesis of cholesterol from acetate
Cholesterol biosynthesis: occurs in the liver
1. Acetyl CoA [C2] + acetoacetyl CoA [C4]  HMG CoA [C6]
2. HMG CoA [C6] + 2NADPH + 2H+  mevalonate [C6] + 2NADP+ + HS-CoA
HMG CoA reductase (3-hydroxy-3-methylglutaryl CoA reductase)
This is the regulated step in cholesterol synthesis: it is inhibited by statins, mevalonate, bile acid and
cholesterol
3. Isoprenoid metabolism (head-to-tail condensations of isoprene units): mevalonate  squalene
a.
Mevalonate [C6]  isoprene units [C5]
b.
C5 + C5  C10
c.
C10 + C5  C15 (farnesyl pyrophosphate)
There is a branched pathway at farnesyl pyrophosphate
d.
C15 + C15  C30 (squalene)
4. Cyclisation reaction: squalene  cholesterol
a.
Squalene [C30]  lanosterol [C30]
b.
Lanosterol [C30]  cholesterol [C27]
Outline the synthesis of bile acids and steroid hormones from cholesterol
Synthesis of steroid hormones:

Site of synthesis: gonads and adrenal glands

5 classes: progestins, glucocorticoids, mineralocorticoids, androgens and oestrogens

Pregnenolone is the precursor of all steroid hormones:
Cholesterol [C27]  pregnenolone [C21]
Cholesterol desmolase: catalyses side-chain cleavage of cholesterol

Regulation: the level of steroid is controlled by their rate of synthesis
Synthesis of bile acids:

Function of bile salts: facilitate digestion via emulsification of dietary fats

Site of synthesis: liver

Enterohepatic circulation: circulation of bile between the liver (site of synthesis) and the small
intestine (site of action)

Regulated step of bile acid synthesis:
Cholesterol  7-α-hydroxycholesterol
Cholesterol 7-α-hydroxylase: this is a cytochrome P450 monooxygenase
Describe the mechanism of transport of cholesterol around the body and its uptake into cells
Fat digestion in the GI tract:
1.
Large fat globules are emulsified by bile salts in the duodenum
2.
Lipase (a pancreatic enzyme) digests fat  fatty acids + monoglycerides
3.
Fatty acids and monoglycerides form micelles, along with bile salts
4.
Fatty acids and monoglycerides leave the micelles and diffuse into epithelial cells
5.
Chylomicrons containing fatty substances are transported out of epithelial cells into lacteals
6.
Fatty substances are carried away from lacteals by lymph
Lipoproteins and cholesterol transport:

Cholesterol is insoluble in blood; it is transported in the circulatory system within lipoproteins

Structure of a plasma lipoprotein: a core of cholesterol esters and/or triacylglycerols surrounded by a
shell of cholesterol, phospholipids and proteins (apoproteins)

Apoproteins: (i) emulsify lipids and (ii) contain cell-targeting signals (determine which cells cholesterol
is transported to)

Lipoproteins (in order of increasing density/decreasing size): chylomicrons; VLDL; LDL; IDL; HDL

Lipoproteins can be separated either by ultracentrifugation or by electrophoresis

Chylomicrons, VLDL and LDL: transport cholesterol to peripheral tissues
Mechanism of cholesterol transport:
1.
Chylomicrons transport triacylglycerols and cholesterol from the intestine to the liver
Triacylglycerols (in chylomicrons) are hydrolysed by lipoprotein lipase in the capillaries
2.
Chylomicron remnants and cholesterol are taken up by the liver, where they are repackaged as VLDLs
3.
VLDLs transport excess triacylglycerols and cholesterol from the liver in the bloodstream
Triacylglycerols (in VLDLs) are hydrolysed by lipoprotein lipase in the capillaries: VLDLs  IDLs
4.
IDLs are either converted into LDLs by lipoprotein lipase or they are taken up by the liver
5.
LDLs transport cholesterol to peripheral tissues or they are taken up by the liver
6.
HDLs transport cholesterol from peripheral tissues to the liver or to endocrine glands for steroid
hormone synthesis
Draw a diagram of low density lipoprotein (LDL)
LDL structure:

Core: cholesterol in the form of cholesteryl esters

Shell: unesterified cholesterol, phospholipids and a single apolipoprotein B-100 molecule
Apolipoprotein B-100 (Apo B-100):

Maintains the solubility of LDLs in the aqueous environment of the blood

Acts as a recognition site and binds with the receptor of target cells
Explain why disturbances in cholesterol homeostasis cause disease
Cholesterol homeostasis:

Dietary cholesterol reduces the activity of HMG CoA reductase

Hepatocytes (and enterocytes): synthesise cholesterol de novo

All cells except those of the liver and intestine obtain cholesterol from the plasma
LDLs are the primary source of cholesterol


Receptor-mediated endocytosis: LDL uptake
o
LDLs bind to LDL receptors on the surface of cells and are taken up by endocytosis
o
LDL cholesterol is removed from the blood
o
The mevalonate pathway (and hence cholesterol biosynthesis) is suppressed
When cholesterol is abundant inside the cell, new LDL receptors are not synthesised; therefore uptake
of additional cholesterol from plasma LDLs is blocked
Cholesterol homeostasis dysfunction:

LDLs transport cholesterol via the arteries

LDLs may be retained in the arteries and form atherosclerotic plaques

Increased LDL cholesterol levels are associated with: atherosclerosis and cardiovascular disease
Hypercholesterolemia:

Caused by high serum LDL cholesterol levels (high LDL to HDL ratio)

Hypercholesterolemia induces the formation of atherosclerotic plaques in arteries (as above)
Familial hypercholesterolemia: hereditary form of hypercholesterolemia

Cause: there is an absence/deficiency of functional LDL receptors due to a mutation in the LDL
receptor gene

Cholesterol is deposited in various tissues due to the high serum level of LDL cholesterol

Serum cholesterol level in homozygotes ~ 680 mg/dl

Serum cholesterol level in heterozygote ~ 300 mg/dl

Desirable serum cholesterol level < 200 mg/dl
Give an example of how a selective enzyme inhibitor can be used as a pharmacological agent
controlling cholesterol metabolism
HMG CoA reductase inhibitors (statins): inhibit the de novo synthesis of cholesterol in hepatocytes

HMG CoA reductase catalyses the formation of mevalonate: this is the committed step in cholesterol
synthesis

Mode of action: statins act as competitive inhibitors of HMG CoA reductase; therefore cholesterol
synthesis in hepatocytes is reduced

The decrease in cholesterol synthesis is detected and LDL receptor production is increased

Therefore more LDL cholesterol is taken up into hepatocytes by endocytosis
Bile acid sequestrants (e.g. cholestyramine): inhibit the intestinal reabsorption of bile salts

Bile salts promote the absorption of dietary cholesterol and dietary fats

Mode of action: bile acid sequestrants bind negatively charged bile salts and prevent their
reabsorption
Control of hypercholesterolaemia:

Dietary restriction of fats and cholesterol

Increased secretion of bile acid sequestrants

HMG CoA reductase inhibitors inhibit cholesterol synthesis
Metabolism 8: Membrane Trafficking
Explain the terms "endocytosis" and "exocytosis"
Endocytosis: uptake of extracellular material into a cell via invagination of the plasma membrane to form
vesicles

Phagocytosis: uptake of solid particles
A large internal phagosome is formed around the extracellular material by the plasma membrane

Pinocytosis: uptake of extracellular fluid and dissolved solutes
Small vesicles are formed around the extracellular material by the plasma membrane

Receptor-mediated endocytosis: molecule-specific endocytosis
A specific receptor on the plasma membrane binds to an extracellular molecule which it recognises
Exocytosis: discharge of intracellular material from a cell via fusion of secretory vesicles with the plasma
membrane
Describe the pathway and cellular locations for synthesis, post-translational modification and
exocytosis of a secreted protein
3 types of intracellular transport:
1.
Gated transport: e.g. nuclear import
2.
Transmembrane transport: e.g. import of newly synthesised proteins into the ER
3.
Vesicular transport: e.g. transport between organelles
Protein targeting: newly synthesised proteins are delivered to the appropriate cellular destinations
Following protein synthesis proteins may be directed to:

A cell membrane: either the plasma membrane or an intracellular (organelle) membrane

The aqueous interior of an organelle

The cell exterior (via secretion)
Secretory pathway: a sequence in which proteins are moved out of a cell
Ribosomes  ER  Golgi apparatus  secretory vesicles  plasma membrane  cell exterior
Protein synthesis: occurs on the ribosomes in the cytosol

A common pool of ribosomes in the cytosol synthesises all proteins: i.e. both proteins which remain in
the cytosol and those which are transported into the ER

The ER signal peptide directs an engaged ribosome to the ER membrane
Signal peptide: the amino acid sequence within a protein which directs its post-translational transport

Polyribosome: the mRNA molecule may remain permanently bound to the ER membrane
Ribosomes move along the mRNA and are recycled

At the end of protein synthesis, ribosomes are released from mRNA and rejoin the common pool of
ribosomes in the cytosol
Translocation of secretory proteins across the ER membrane:
1.
The ER signal peptide at the N-terminus of a protein directs it to the ER membrane
2.
The protein is guided to the ribosome by a signal recognition particle (SRP) which binds to the signal
peptide
3.
The SRP in turn binds to an SRP receptor on the ER membrane: this targets the protein to the ER
4.
Cotranslational translocation: the protein is fed into the ER lumen via a translocation channel while it
is still being synthesised on the ribosome
5.
Post-translational translocation: the protein is fed into the ER lumen after being fully synthesised by
the ribosome
6.
Following translocation of the protein into the ER lumen, a signal peptidase cleaves the ER signal
peptide from the protein
Post-translational modification and quality control:
Post-translational modification: occurs in the ER, Golgi and secretory vesicles

Glycosylation: addition of carbohydrate chains to specific amino acids (ER) and processing (Golgi)

Formation of disulphide bonds (ER): occurs between cysteine residues

Folding of proteins (ER): facilitated by chaperones and ER proteins

Assembly of multimeric proteins (ER): facilitated by chaperones and ER proteins

Specific proteolytic cleavages (ER, Golgi and secretory vesicles)
Quality control: unfolded or misfolded proteins are retained in the ER and transported back into the
cytosol for degradation
Exocytosis:
Soluble and membrane proteins move to their final destinations via the secretory pathway

This is mediated by transport vesicles which transport cargo proteins between organelles

The transport vesicles bud from one organelle and fuse with another organelle
Mechanism of the secretory pathway:
1. Secretory proteins are packaged into anterograde transport vesicles by budding from the ER
2. Anterograde ER transport vesicles fuse together to form new cis-Golgi cisternae
3. ER-resident proteins are retrieved from the cis-Golgi via retrograde transport vesicles
4. Cisternal progression: cis-Golgi cisternae successively move from the cis to the trans face of the Golgi
complex
Cis-Golgi cisternae  medial-Golgi cisternae  trans-Golgi cisternae
5. Golgi-resident proteins are retrieved from later Golgi cisternae (i.e. trans-Golgi cisternae) via
retrograde transport vesicles
6. Vesicles eventually reach the trans-Golgi network: this is a major branchpoint in the secretory pathway
and results in…
a. Constitutive secretion
b. Regulated secretion
c. Lysosome formation: soluble proteins within vesicles bud from the trans-Golgi network and move
to the late endosome and finally to the lysosome
Distinguish "constitutive" and "regulated" secretion
Constitutive secretion: soluble proteins are continuously secreted from the cell

No external signals are required for secretion

Secretion is not affected by environmental factors

The Golgi apparatus is scattered throughout the cytoplasm

E.g. fibroblasts (secrete collagen) and activated B lymphocytes (secrete antibodies)
Regulated secretion: soluble proteins are stored inside the cell

A specific signal is necessary for secretion (neural or hormonal stimulation)

Cells are usually apical or polarised

The Golgi apparatus is between the nucleus and the secretory surface

E.g. goblet cells (secrete mucus) and beta cells of the pancreas (secrete insulin)
Describe the process of receptor-mediated endocytosis and the roles played by endocytic vesicles,
early endosomes, late endosomes, and lysosomes
Receptor-mediated endocytosis: the process whereby cells internalise extracellular material

LDL is taken up into cells via receptor-mediated endocytosis: LDL receptors are localised in clathrincoated pits
Mechanism of endocytosis for internalising LDL:

LDL receptors on the cell surface bind to Apo B-100 on the surface of LDLs

Clathrin-coated pits containing LDL receptor-LDL complexes bud off from the plasma membrane
enclosed in endocytic vesicles (early endosomes)

The vesicle coat is shed and the uncoated early endosome fuses with a late endosome

The late endosome fuses with a lysosome: the constituent lipids and proteins of LDL are hydrolysed
by lysosomal enzymes; cholesteryl esters are hydrolysed by a lysosomal lipase

LDL receptors are recycled to the cell surface
Give a general description of the molecular mechanisms of vesicular transport within cells
Give examples of diseases resulting from defects in the secretory and endocytic pathways
I-cell disease (inclusion cell disease): lysosomal storage defect

Mannose is phosphorylated to mannose 6-phosphate (M6P) by a phosphotransferase at the cis-Golgi
network

The M6P residue acts as a lysosomal sorting signal at the trans-Golgi network

I-cell disease: there is a mutation in the phosphotransferase which catalyses the formation of M6P;
therefore affected individuals lack the M6P sorting signal

Consequences:
o
The lysosomal enzymes are secreted rather than being sorted into lysosomes
o
Undigested glycolipids and extracellular components accumulate in lysosomes (they would
usually be degraded by lysosomal enzymes)
Familial hypercholesterolemia: endocytic pathway defect

The affected individual produces a mutant form of the LDL receptor

This causes impaired endocytosis of LDL which results in high serum LDL cholesterol levels
Metabolism 9: Integration of Metabolism
Outline general features of metabolic activity in liver, brain, muscle, adipose tissue
Metabolism: the sum of all processes in the body
Methods of measuring metabolism:

O2 uptake

CO2 release

Heat production
Metabolic features of tissues:
% Body mass
% Resting MR
(metabolic rate)
Liver
2.5%
Muscle
Heart
Tissue
Metabolites
Other features
20%
Carbohydrates, fats and
amino acids
Glycogen store
40%
Variable
Carbohydrates and fats
1%
10%
Carbohydrates and fats
Brain and
nervous
2%
20%
Glucose (and ketone
bodies during starvation)
Adipose
tissue
15%
Low
n/a
Major source of blood glucose
May have periods of very high
ATP requirement during
vigorous muscle activity
Cannot metabolise fats
Continuously high ATP
requirement
Long term store of fats
Further metabolic features of the liver:
The liver plays a central role in the coordination of metabolism throughout the body:

It receives nutrients absorbed at the intestines immediately (via the hepatic portal vein)

Performs a wide repertoire of metabolic processes including:
o
Glycolysis
o
Glucose production (gluconeogenesis)
o
Glucose storage

High metabolic activity: 2.5% of body mass vs. 20% of resting metabolic rate

It can interconvert nutrient types

Plays a central role in the maintenance of blood glucose at 4.0-5.5 mM

It performs lipoprotein metabolism (transport of triglycerides and cholesterol)
Gluconeogenesis: the process whereby glucose or glycogen is made from oxaloacetate

Primarily occurs in the liver (and also in the kidneys to a small extent)

Requires ATP hydrolysis: 6 ATPs are used

Certain enzymes bypass the irreversible steps of glycolysis

Gluconeogenesis and glycolysis are reciprocally regulated (i.e. they inhibit each other)
Gluconeogenesis pathway: (only the enzymes for the bypass reactions are shown)
1.
Pyruvate  oxaloacetate
Pyruvate carboxylase (in the mitochondria)
2.
Oxaloacetate  phosphoenolpyruvate
Phosphoenolpyruvate carboxykinase (in the cytosol)
3.
(Phosphoenolpyruvate … fructose 1,6-bisphosphate)
4.
Fructose 1,6-bisphosphate  fructose 6-phosphate
Fructose 1,6-bisphosphatase
5.
(Fructose 6-phosphate  glucose 6-phosphate)
6.
Glucose 6-phosphate  glucose
Glucose 6-phosphatase: specifically found in the liver
Muscle
Liver
Enzymes
Hexokinase I
Hexokinase IV and glucose 6-phosphatase
Reaction
Glucose  glucose 6-phosphate
Glucose  glucose 6-phosphate
High glucose affinity: the rate is half-maximal at
0.1 mM
Low glucose affinity: the rate is half-maximal at
4 mM
Highly sensitive to G6P inhibition
Less sensitive to G6P inhibition
Saturated at low [glucose]
Small ∆[glucose]  big ∆rate
Therefore it is poor for glucose regulation
Therefore it is good for glucose regulation
Hexokinase
Metabolism of ethanol: CH3CH2OH  CH3CHO  CH3COOH  CH3CO-CoA

Ethanol metabolism is unregulated; therefore this leads to perturbation of normal metabolism

Ethanol + NAD+  acetaldehyde + NADH + H+
Alcohol dehydrogenase

Medicinal drugs metabolised by a similar pathway may cause liver damage in alcoholics

Unregulated ethanol metabolism leads to the accumulation of NADH
Glycolysis is suppressed due to a lack of NAD+

TCA intermediates are reduced to sustain ethanol metabolism:
Lactate/malate + NAD+  pyruvate/oxaloacetate + NADH + H+

Lactate accumulates as it cannot be oxidised to pyruvate due to a lack of NAD+
Regulation of glucose metabolism:
Glucose metabolism is regulated at a step which is:

Strongly dependent on enzyme activity

Unique to the pathway

Early in the pathway
Regulatory mechanisms include:

End-product inhibition: i.e. the products of the reaction pathway regulate glucose metabolism

Signalling molecules which relay information from other pathways or cells
Give four examples of blood-borne hormones which act as metabolic regulators
Insulin
Glucagon
Secreted by the islets of Langerhans of the pancreas
Secreted in response to a rise in blood glucose level
Secreted in response to a fall in blood glucose level
Stimulates uptake and use of glucose
Stimulates production of glucose by gluconeogenesis
Stimulates storage of glucose as glycogen and fat
Stimulates breakdown of glycogen and fat  glucose
Adrenaline
Glucocorticoids
Secreted by the adrenal glands
Strong and fast metabolic affects to mobilise glucose in
response to ‘fight or flight’ stimuli
Steroid hormones which increase the synthesis of
metabolic enzymes that catalyse formation of glucose
Know the effects of eating and fasting on metabolism
In a real person, metabolism has to be able to cope with:

Different types of food

Sporadic food intake

The different requirements of different tissues
After a meal: blood glucose rises initially and then falls
Control mechanisms for the initial rise in blood glucose:

Islets: increased insulin secretion (and reduced glucagon secretion)

Liver: increased glucose uptake; glucose is used for glycogen synthesis and for glycolysis
Acetyl CoA produced by glycolysis is used for fatty acid synthesis

Muscles: increased glucose uptake; glucose is used for glycogen synthesis

Adipose tissue: increased triglyceride synthesis

Increased usage of metabolic intermediates throughout the body
Summary of effects due to the rise in blood glucose:
1.
High glucose stimulates insulin release from the pancreas
2.
High insulin stimulates glucose uptake into the liver and tissues
3.
Insulin stimulates anabolic pathways
Control mechanisms for the subsequent fall in blood glucose:

Islets: increased glucagon secretion (and reduced insulin secretion)

Liver: glucose production occurs due to gluconeogenesis and glycogen breakdown

Fatty acid breakdown: this provides an alternative metabolite for ATP production
This is important for preserving glucose for the brain

Glucagon stimulates catabolic pathways
Adrenaline:
Adrenaline has similar effects to glucagon on the liver
It also has additional effects on:

Skeletal muscle: adrenaline stimulates glycogen breakdown and glycolysis

Adipose tissue: adrenaline stimulates fat lipolysis (hydrolysis) to provide other tissues (i.e. tissues
other than the brain) with alternative metabolites to glucose (i.e. fatty acids)
After prolonged fasting (i.e. longer than can be covered by glycogen reserves):

The glucagon to insulin ratio increases further

Adipose tissue: hydrolysis of triglyceride occurs to provide fatty acids for metabolism

The quantity of TCA cycle intermediates decreases to provide substrates for gluconeogenesis

Protein breakdown occurs: this provides amino acid substrates for gluconeogenesis

Liver: ketone bodies are produced from fatty acids and amino acids to partially substitute the glucose
requirement of the brain
Fatty acid breakdown:
1.
ß-oxidation: fatty acids  acetoacetyl CoA
Branched pathway at acetoacetyl CoA:
2.
Acetoacetyl CoA  acetyl CoA/acetate
Branched pathway at acetate:
3.
Acetate  acetone/ß-hydroxybutyrate
Acetate, acetone and ß-hydroxybutyrate are ketone bodies
There is excess accumulation of acetyl CoA:

It is used by the heart under normal circumstances

It is used by the brain when glucose availability is low
Brain:

Glucose dependent metabolism: the brain requires a continuous supply of glucose for metabolism
o
Hypoglycaemia: causes faintness and coma
o
Hyperglycaemia: may cause irreversible damage

The brain cannot metabolise fatty acids directly; it can metabolise ketone bodies

Ketone bodies (ß-hydroxybutyrate) can partially substitute for glucose
Describe glucose interactions with lipid and amino acid synthesis and breakdown
See flow charts
Know basic details of contractile metabolism in muscle
Skeletal muscle:
During light contraction: ATP demand is met by oxidative phosphorylation using O2 and blood-borne
glucose and fatty acids
During vigorous contraction: ATP demand cannot be met by oxidative phosphorylation since the diffusion
of O2 and blood-borne metabolites is a limiting factor

Glycogen stores in the muscle are broken down to produce ATP

Under anaerobic conditions, pyruvate is converted into lactate and H+ which can leave muscle
Aerobic exercise:
1.
Actomyosin ATPase, Ca2+ ATPase and Na+–K+ cause muscle contraction
2.
Contractions stimulate:
3.
o
Increased ATP demand
o
Increased glucose transport from the liver to muscles via the bloodstream
Adrenaline stimulates:
o
Increased muscle glycolysis: this produces ATP
o
Increased gluconeogenesis
o
Increased fatty acid production
Anaerobic exercise:
1.
ATP demand cannot be met by O2 diffusion
2.
Transport cannot keep up with the demand for glucose
3.
Breakdown of glycogen stores in the muscle increases
4.
Lactate production increases: it is transported to the liver via the bloodstream
5.
Recovery: the liver uses lactate to form glucose
Heart:

The heart must beat constantly; therefore it is designed for completely aerobic metabolism as it has a
large number of mitochondria

It can use TCA cycle substrates for metabolism: e.g. fatty acids and ketone bodies

Loss of oxygen is devastating: it leads to cell death and myocardial infarction since energy demand is
much greater than energy supply
Be able to describe some of the metabolic disturbances that arise in diabetes
2 types of diabetes:
1.
Type I diabetics cannot produce insulin
2.
Type II diabetics have reduced responsiveness to insulin
Metabolism is controlled as if for starvation, regardless of dietary uptake
Complications of diabetes:

Hyperglycaemia with progressive tissue damage

Increase in plasma fatty acids and lipoproteins with possible cardiovascular complications

Increase in ketone bodies with possible acidosis

Hypoglycaemia with constant coma if the insulin dosage is imperfectly controlled