2016 - BIOL1007 Lectures - Colour PDF

BIOL1007 Notes - 2016
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Metabolism Module
Simon Worrall
School of Chemistry and Molecular
Biosciences
© Simon Worrall 2016
1
Section 1
An Overview of Metabolism
Learning Objectives
1. Understand what constitutes metabolism
2. Appreciate that there are different types of
metabolic pathways
3. Recognize that
 Metabolism is dynamic
 Catabolic pathways are primarily about energy
production
 Energy is released by oxidation
 Oxidation is all about loss of electrons
2
Overview
• Metabolism is the sum of chemical reactions,
usually catalysed by enzymes that occurs in
the living cell ‐ the total of both anabolic and
catabolic reactions
• Metabolism is highly regulated in various ways
• Chemical reactions in the cell occur as
‘pathways’ i.e. a particular sequence of
chemical reactions.
• Each of the consecutive steps in a pathway
produces a chemical product (metabolite or
intermediate) which is different from its
reactants.
• There is evidence that enzymes which catalyse
a specific sequence of chemical reactions (a
pathway) may function as highly organised
multienzyme complexes (c.f. Luke Guddat lectures).
• As a consequence of these complexes each
metabolite seems to ’channelled’ directly from
the active site of one enzyme to the active site
of the enzyme catalysing the next step in the
pathway. 6
3
Pathways Can Be Organised Differently
Linear Branched
Cyclic Complex
Batmanian et al. Figs 14‐1 to 14‐4 7
Two Types of Pathways
Anabolic pathways Catabolic pathways
• Biosynthetic (make molecules) • degradative (break molecules)
• reductive (require electrons) • oxidative (donate electrons)
• commonly use NADPH as an • use NAD+ and FAD as electron
electron donor carriers (acceptors)
• energy requiring • yield energy
• pathway is usually divergent • pathway is usually convergent
4
All is in a State of Flux
Catabolic and anabolic pathways proceed at
same time in a dynamic steady state. The
energy‐yielding catabolic pathways are
balanced by the anabolic pathways.
Dynamic Steady State for B
r1 r2
A B C
synthesis degradation
If rate of synthesis (r1) = rate of degradation (r2) then [B]
remains constant
9
Anabolic vs. Catabolic
• Anabolic pathways are not usually the reverse
of catabolic pathways.
• Corresponding anabolic and catabolic
pathways may share some common reversible
reactions but are controlled by different
regulatory enzymes catalysing irreversible
reactions.
• Energy‐requiring anabolic reactions are
coupled to energy yielding catabolic pathways
– overall G for pathway must be –ve.
10
5
Comprehensive
Metabolic Map
11
Don’t Miss the Wood for the Trees!
Metabolism doesn’t have to be complicated
Pathways reduce to concepts
Concepts help understand what goes on in
cellular metabolism for life
• “flows” – inputs, outputs & passage along pathways
• “pools” – amounts of molecules that are filled
(supplied) or emptied (by demand) by catabolic and
anabolic pathways (substrate pool, energy pool,
electron pool, nutrient pools)
• “space & time” – not all pathways are necessarily
occurring at the same time, in the same place
12
6
We are at this Level!
How Cells get Energy
Glycolysis
Electron
transport chain
and oxidative
phosphorylation
The TCA
cycle
13
Metabolic Concept Map
ENERGY TRANSFER
MOLECULES
Lehninger 3rd Ed, p 487.

14
7
Metabolism Needs to be Controlled
Metabolism needs to be regulated
(controlled)
 to prevent futile (substrate) cycles
 to respond to physiological needs
 to respond to changes in energy demand
15
Controlling Metabolism
Metabolism regulated by:
• thermodynamics of reactions
 influence of substrate concentration (mass action effect)
• amount of enzymes ‐ increase or decrease rate
of synthesis
• activity of enzymes
 allosteric ‐ substrate feedback mechanisms
 covalent modification e.g. phosphorylation
• compartmentalisation
 keeping related reactions apart by putting them in different
locations (organs, tissues, organelles).
16
8
Obtaining Energy from Food
Food is oxidised ‐ by chemical reactions that release energy
 What is an oxidation reaction?
 How do these reactions release energy?
This energy is converted into other forms that can be used in
energy‐requiring reactions in the cells of the body
Most commonly, the energy is used to synthesise adenosine
triphosphate (ATP) ‐ a “high energy” compound that can be
used to provide energy to drive chemical reactions that
otherwise would not occur.
 What is ATP?
 How is it made?
 How is it used to provide energy?
17
Stages of Fuel Oxidation
STAGE 1
O2 CO2 Carry usable energy as
electrons
Metabolic Reduced coenzymes
fuel (NADH + H+, FADH2)
STAGE 2
O2 H2O
Reduced coenzymes ATP
(NADH + H+, FADH2)
Carry usable energy as High‐energy
electrons phosphate
18
9
Fuel Sources
• Macronutrients
catabolised to common
intermediate, acetyl CoA
• Acetyl‐CoA oxidised in
TCA cycle producing
NADH & FADH2
• Energy derived from
reoxidised NADH and
FADH2 drives ATP
synthesis
• Remember – all is in
balance ‐ homeostasis
19
Oxidation of Food Releases Energy
Oxidation Reactions
Food + O2 CO2 + H2O

Catabolism
Energy
Oxidative phosphorylation by
electron transport chain (ETC)
Adenosine Adenosine
diphosphate triphosphate
(ADP) + Pi (ATP)
20
10
Oxidation‐reduction Reactions
In an oxidation reaction,
electrons are lost.
In a reduction reaction,
electrons are gained.
Redox reactions are reactions Oxidation Involves Loss – Reduction Involves Gain

in which electrons are passed
from a reduced donor to an
oxidised acceptor
Donor‐e‐ + Acceptor ö Donor + Acceptor‐e‐

Fe2+ + Cu2+ ö Fe3+ + Cu+
Reductant Oxidant Oxidised Reduced
21
Oxidation of C Atoms in
Biological Systems ‐ 1
Covalent bond – shared electrons (one from each atom)
Distribution not equal because C atom is more electronegative
than H
Both electrons spend more time around the C nucleus than
around the H nucleus.
So, the C atom has effectively 2 electrons, the H has none.
C + H C H
22
11
H is replaced by another C
 Balanced e‐ distribution
 Original C effectively loses an electron and is
oxidised.
C H C C
23
Second C is replaced by O
 Oxygen is more electronegative than carbon and
therefore attracts electrons more strongly.
– Original C effectively loses the second electron
and is further oxidised.
C C C O
24
12
Food molecules are rich in C‐H or C‐C bonds, with
relatively few C‐O bonds
 The C atoms in food molecules are therefore in a reduced
state
In metabolism, C‐H and C‐C bonds become C‐O (CO2)
 The food molecules are therefore oxidised.
 e.g. fatty acids react with O2 to form CO2 (& H2O)
‐OOC‐(CH ) ‐CH
2 n 3 + O2 ö CO2 + H2O
25
 Oxidation of glucose also yields energy
26
26
13
Biological Redox Reactions often
use NAD+
Many oxidation reactions in the cell involve the transfer
of 2 e‐ in the form of H‐ (hydride) ions to nicotinamide
adenine dinucleotide ions (NAD+).
NAD+ is therefore reduced to NADH.
 The reduction of NAD+ is thermodynamically unfavourable
 Requires an input of energy, derived from an oxidation
reaction
Reoxidation of NADH releases energy
FAD/FADH2 also used
 FAD accepts 2 electrons as hydrogen atoms.
27
Things to Think About
• Metabolism is an exercise in thinking about
POPULATIONS of components (there are many
thousands of individual molecules of substrate,
product and enzyme).
• All is in a state of flux.
• Metabolic pathways are a concept to aid our
understanding – with a few exceptions there is no
strict spatial organisation of one enzyme with
respect to another.
28
14
• Energy (especially Gibb's free energy, G) ‐ how
products of a reaction are in a lower energy state;
that the difference in the energy state between
product and substrate can be large or small and
how that influences a reaction. Principle applies
to overall pathways.
• Catabolic pathways yield energy, generally by
oxidation.
• We can think about intermediary metabolism in
terms of electron flow yielding useful energy
• How might we use this energy and it what form?
29
Section 2
Glycolysis – the Oxidation of
Glucose
30
15
Learning Objectives
1. Appreciate the central importance of
glycolysis in intermediary metabolism
 that It is 1st stage of metabolic respiration.
2. Recognize that glucose is split to 2
molecules of 3‐carbon pyruvate
 that little energy is generated as ATP or
potential energy as reducing equivalents.
3. Understand that redox balance must be
maintained
 that the process needs to be regulated.
31
Glucose Oxidation
Energy can be released
explosively BUT we need to
obtain the energy in a
controlled manner
Flour is highly explosive
when presented with an
ignition source in an O2
Starch, a polymer of glucose. atmosphere
Main constituent of flour
Uncontrolled oxidation! 32
16
BIOL1007: Metabolism Module Glucose Oxidation ‐ Animation
https://www.youtube.com/watch?v=X‐ZZETT6F‐s
33
Glucose Oxidation Occurs in Stages
34
17
Key Points about Glycolysis
Glucose
(6C)
NADH
GLYCOLYSIS Oxidative
phosphorylation
ATP
2Pyruvate
(3C) Oxidative
phosphorylation
2 Acetyl CoA (2C) NADH
CO2 35
Oxidation of Glucose to Pyruvate
The sequence of reactions that converts 6‐carbon
glucose to the 3‐carbon metabolite pyruvate is
called glycolysis.
Pyruvate (or pyruvic acid) is a key compound of
central importance in metabolism
 it has 3 carbon atoms carboxylic
 it is an "‐keto acid" acid
group
‐keto group
(also 2‐oxo)
36
18
Glycolysis in Overview ‐ 1
• Glycolysis is the first
step in the production
of energy from sugars
• Sugars are highly
reduced. Their
OXIDATIVE breakdown
will produce energy.
• Glycolysis is linked to
energy production.
• Pyruvate oxidation
supplies acetyl CoA to
the TCA cycle.
• The TCA cycle is linked
with oxidative Campbell et al Fig 9.6
phosphorylation which
produces ATP.
37
Glycolysis in Overview ‐ 2
Glycolysis has two phases
 energy investment
 energy generation
Campbell et al Fig 9.8 38
19
BIOL1007: Metabolism Module Glycolysis Animation
https://youtu.be/3GTjQTqUuOw
39
Glycolysis has 3 Functional Parts ‐ 1
Lehninger Fig 14.2
What is achieved?
 Glucose is trapped inside the cell
 Glucose is converted for cleavage into 2 x 3‐carbon units
 2 ATP consumed to drive unfavourable reactions
 No oxidation has occurred
40
20
What is achieved?
 6‐C molecule is cleaved
into 2 non‐identical 3‐C
fragments
 The 3‐C fragments are
interconverted by
isomerization 4
 No ATP is used or
produced
 No oxidation has
occurred
Lehninger Fig 14.6
41
What is achieved?
 The 3‐C fragment
glyceraldehyde 3‐
phosphate is
oxidized and
NADH is produced
 ATP is synthesized
in two separate
"substrate‐level
phosphorylation"
steps
Note that, per glucose molecule, all
intermediates in this third part of the
pathway are multiplied by 2. Lehninger Fig 14.X
42
21
Regulation of Glycolysis ‐ 1
• Glycolysis is regulated.
• Activity is increased in the fed state (activated by
insulin).
• Promotes storage of “food energy”
• Activity is decreased in the fasting state (inhibited
by glucagon).
• Promotes use of stores for energy.
• Activity is increased when there is an ATP demand
e.g. exercise.
• Irreversible reactions are regulatory steps.
43
Insulin activates the three
enzymes that catalyse the
irreversible steps of
glycolysis.
Glucagon inhibits these
enzymes‐ glycolysis is
inhibited in the fasting state.
These hormones act through
2nd messenger systems.
Remember principles of
metabolic regulation –
regulate irreversible steps
44
22
In addition to regulation by
insulin and glucagon,
glycolysis is regulated by
ATP demand.
PFK is inhibited by ATP and
activated by ADP/AMP.
PFK is also regulated by
citrate (signals CAC has
sufficient substrate).
Remember principles of
enzyme regulation lectures
Campbell et al Fig 9.20 45
Summary of Glycolysis
• Oxidation of glucose via the
glycolytic pathway results in
the generation of 2
molecules of pyruvate.
• Glycolysis has used two
molecules of NAD+
• Need to regenerate NAD+ in
order to keep glycolysis
going.
• Ability to achieve this
depends upon the use of
pyruvate e.g. fermentation.
46
23
Summary of Glycolysis
• This sequence of reactions is
very similar in all organisms &
in many different types of cell.
• The fate of pyruvate depends
on the type of cell and the
conditions.
• Little ATP or potential ATP in
the form of reduced electron
carriers has yet been
produced.
• Further energy production
depends on the fate of
pyruvate.
Berg et al Fig 16.10
47
Anaerobic Glycolysis
In anaerobic conditions, the electron transport chain
will not operate ‐ needs O2 as the final electron
acceptor.
Glycolysis produces 2 NADH
Glucose
Without O2, cannot reoxidise
NADH via Electron Transport
DHAP G3P + Pi + NAD+ Chain.
Need another way of reoxidising
1,3BPG + NADH NADH or glycolysis will stop ‐
supplies of NAD+ are very
Pyruvate limited.
48
24
BIOL1007: Metabolism Module Maintaining Redox Balance
Berg et al. Fig 16.10
Pyruvate can be converted to:
• Ethanol ‐ fermentation in yeast in anaerobic conditions
• Lactate ‐ reduction in anaerobic conditions in muscle
• Acetyl CoA ‐ oxidation in aerobic conditions
Subsequent metabolism of pyruvate regenerates NAD+
49
Aerobic Fate of Pyruvate

Pyruvate feeds into TCA cycle
Glycolysis and TCA occur in different compartments
 Glycolysis – cytosol
 TCA – mitochondrion
Pyruvate transported inwards
 Pyr‐H+ symporter
Pyruvate dehydrogenase
 Multi‐enzyme complex
 Contains thiamine pyrophosphate – derived from vitamin B1
 Arsenic poisoning
50
25
Aerobic Glycolysis
Acetyl CoA production
Mitochondrial
Cytosol matrix
Transporter
Section 16.1 in Lehninger
51
Central Role of Acetyl CoA

• Central role in
metabolism
• End product of
carbohydrate, fat &
amino acid
breakdown
• Substrate for the
TCA cycle
• Rich source of NADH
for the electron
transport chain
Fig 16.1
52
26
BIOL1007: Metabolism Module …… and of the TCA Cycle
Batmanian et al 16‐01
53
• Why do we have so many steps in this process of
glycolysis?
• Why are some steps reversible and some not?
• Pyruvate is the end product. What determines
how this is used?
• Are these all or none options for the use of
pyruvate?
54
27
Section 3
The Tricarboxylic Acid Cycle
55
Learning Objectives
1. Understand the pivotal role of TCA cycle in
substrate oxidation.
2. Recognize the importance of reducing
equivalents in energy flow.
3. Recognize that the TCA cycle also plays
other roles.
4. Understand the importance of regulation
and the linkage of glycolysis to the TCA
cycle via acetyl CoA.
56
28
The Tricarboxylic Acid Cycle

• Also known as the
Citric Acid Cycle or
Krebs’ cycle
• The TCA cycle is the
final common
pathway for the
oxidation of fuel
molecules
• AMPHIBOLIC
• May also provide
small building blocks
(4C, 5C, 6C) for
synthesis of larger
molecules. Batmanian et al Fig 16‐01
57
TCA Cycle – The Movie
www.youtube.com/watch?v=‐cDFYXc9Wko
58
29
BIOL1007: Metabolism Module Outline of TCA Cycle
Batmanian et al Fig 16‐01
59
Biological Redox Reactions often
use NAD+
Many cellular oxidation reactions involve the transfer of
2 e‐ as H‐ (hydride) ions to NAD+.
Consequently NAD+ is reduced to NADH (+ H+).
 The reduction of NAD+ is thermodynamically
unfavourable
 Requires an input of energy, derived from the
oxidation reaction
Reoxidation of NADH releases energy
FAD/FADH2 also used
 FAD accepts 2 electrons as hydrogen atoms.
60
30
What is Achieved in the TCA cycle?
• One 2‐carbon fragment is oxidized to 2 CO2 . (Acetyl CoA)
• Four oxidations occur, generating NADH/FADH2.
• One high energy phosphate anhydride bond is made. (GTP)
• The carrier molecule oxaloacetate is used and regenerated.
Oxaloacetate is a key
Oxaloacetate acts as a compound of central
“carrier”, there is no importance in metabolism.
nett synthesis
(start of gluconeogenesis)
It has 4 carbon atoms.
Remember the
principle of dynamic It is an ‐keto acid.
steady‐state. (precursor for amino acid
aspartate)
61
Energy Production
• Note again that there is little direct substrate
level production of energy (ATP).
 Only one molecule of GTP formed
• Maximal production of ATP requires oxygen and
the reactions of oxidative phosphorylation.
 TCA cycle can’t function without O2
• The reduced flavin and nicotinamide electron
carriers are oxidized in oxidative phosphorylation.
62
31
The TCA Cycle only Operates in
Aerobic Cells
• Although no O is used in the cycle, O2 is required
to re‐oxidize NADH and FADH2
NADH + H+ + 0.5 O2 NAD+ + H2O
FADH2 + 0.5 O2 FAD + H2O via ETC
• These coenzymes are present in limited amounts
in the cell and must be recycled continuously
• The enzymes of the citric acid cycle are located in
the mitochondrion
Remember cytosolic glycolysis can operate under
anaerobic conditions 63
How Much Energy is Produced?
• TCA Cycle
Acetyl‐CoA + 3 NAD+ + FAD + GDP + Pi + 2 H2O 
2 CO2 + 3 NADH + 3H+ + [ADH2]+ GTP + CoA‐SH
G0 = ‐40 kJ/mol
• Combustion of Acetate
Acetate + 2 O2 + H+  2 CO2 + 2 H2O
G0 = ‐850 kJ/mole
• Difference
Therefore, 810 kJ/mole remain in the high energy
products ‐ GTP, NADH and FADH2
This energy will be harvested on oxidative
phosphorylation. 64
32
What the TCA Cycle Produces
The overall reaction catalysed by the cycle produces:
3 NADH = 9 ATP
1 FADH2 = 2 ATP
1 GTP = 1 ATP
Total = 12 ATP per acetyl CoA
BUT ATP is only gained from NADH and FADH2 by

linking TCA cycle to the ETC and oxidative
phosphorylation and assumes highly efficient
conversion (coupling).
65
TCA cycle is also a Source of
Carbon Substrates
The TCA cycle provides oxidized carbon skeletons
for many important biosynthesis pathways
Pyruvate 3C Amino acids (val, leu, ala)
‐Ketoglutarate 5C Amino acids (glu, gln, pro, arg), purines
Succinyl‐CoA 4C Porphyrins (haemoglobin, cytochromes)
Fumarate & Amino acids (asp, asn, thr, met, Ile, lys),
4C
Oxaloacetate pyrimidines
Oxaloacetate 4C PEP (gluconeogenesis), amino acids (phe, tyr, try)
66
33
Regulation of the Cycle ‐ 1
1. Flux rates
 [substrate] stimulates
 [products] inhibit
2. Allosteric control of
PDH by cellular energy
state
 ATP/AMP ratio
3. Enzyme complexes
67
Regulation of the Cycle ‐ 2
Pyruvate dehydrogenase complex
 Inhibited by NADH, Acetyl‐CoA,
ATP
 Activated by CoA‐SH, NAD+
Citrate synthase
 Inhibited by ATP, NADH, Succinyl‐
CoA
Isocitrate dehydrogenase
 inhibited by ATP
 Activated by ADP and NAD+
‐Ketoglutarate dehydrogenase
 Inhibited by NADH, Succinyl‐CoA
 Activated by AMP
All aimed at providing energy when
it is needed. From Batmanian et al. 68
34
• If the TCA is a cyclic process, how do we actually
gain anything?
• Does the speed at which the cycle turns matter,
i.e. the rate of flux through the pathway?
• Does it matter that the pathway is linked to other
pathways – does this present advantages or pose
problems (or both)?
• Have we got any useful energy yet?
69
Section 4
Oxidative Phophorylation
70
35
Learning Objectives
1. Recognize that although aerobic respiration so far
has yielded some energy as ATP this is not much,
most is still locked away in NADH & FADH2.
2. Recognize that we still haven’t regenerated NAD+
so we are at risk of compromising glycolysis.
3. Understand that we accomplish both of the
objectives above in the final stage of respiration
 that this involves two processes: electron transport
coupled to phosphorylation of ADP to ATP.
4. Recognize the importance of O2 as the final
electron acceptor.
71
Cellular Respiration –
Maximising Energy Production
Campbell et al 9.6
72
36
Oxidative Phosphorylation
Located in the inner mitochondrial membrane
 Inner membrane: impermeable to H+.
 Outer membrane: freely permeable to H+.
Electrons pass through ETC protein complexes.
 Passed between complexes by mobile electron carriers ‐
coenzyme Q (ubiquinone) and cytochrome c
H+ 3 H+ H+
H+ H+ H+ H+
H+ H+ H+
H+
H+
H+ H+
V
H+ H+
H+
H+ H+
H+
H+ ADP + Pi ATP H+
H+
H+ H+ H+
2 H+ + ½ O2 + 2 e-  H2O NADH NAD+
H+ H+ H+
H+ IV III I
H+ H+
C UQ
2 H+ 4 H+ H+
4 H+
73
The Electron Transport Chain
NADH is used to drive ATP synthesis Reduction potentials & the ETC
Campbell, Reece, Myers 7th Edn
74
37
Electron Carriers
Cytochromes
 Proteins containing haem
prosthetic groups
 Fe3+/Fe4+ metal centres can transfer
single e‐
Iron‐sulphur Proteins
 contain an Iron‐sulphur cluster
 Fe3+/Fe4+ metal centres can transfer
single e‐
Copper proteins
 Contain an copper ion
 Cu+/Cu2+ metal centres can
transfer single e‐
75
Electron Transport ‐ Animation
www.youtube.com/watch?v=Ak17BWJ3bLg
76
38
Products of Electron Transport
What has this pathway produced?
 ATP? ‐ No
 Other high energy molecules that could power ADP
phosphorylation? ‐ No
What has it produced?
 A proton concentration gradient across mitochondrial
inner membrane generating a “proton motive force”
 A charge gradient from the electrogenic transport of H+
 Two components: Δ pH & Δ ψ (difference in charge)
 Tendency of Δ [H+] toward equilibrium
 Source of energy
77
Chemiosmotic Model
First proposed by Peter Mitchell (against much opposition) in
1960s. Lead to Nobel prize in Chemistry, 1978.
78
39
ATP Synthase is a Rotary Motor
H+ flow through a membrane‐spanning portion of
ATP synthase
 Free energy released
is used to drive
synthesis of ATP from
ADP and Pi.
 Transport of ~3 H+ per
ATP made.
 Acts as a rotary motor.
Thomas M. Terry, UConn (http://nhscience.lonestar.edu/biol//bio1int.htm)
79
Cellular Respiration ‐ the Whole Works!
80
40
• Can aerobic and anaerobic metabolism operate
simultaneously?
• Can increasing oxygen supply, e.g. by breathing
harder, speed up aerobic respiration?
• After a mixed meal (CHO, fat and protein), do all
contribute to energy supply via the mechanisms
we have discussed so far?
81
Section 5
Metabolic Integration
82
41
Learning Objectives
In the 1st lecture we noted the many different
metabolic pathways that exist in living
organisms. These different pathways are
interconnected and need to be regulated and
coordinated.
From this lecture you should:
1. Recognize how muscle uses different fuels
depending on energy demand
2. Understand how the body responds to times
of nutritional need e.g. fasting and
starvation, by altering its fuel metabolism.
83
Why Integrate?
• Not all metabolic pathways are active in all
cells.
• Organs specialise e.g. liver is major fuel
provider, brain is major fuel consumer.
• Organs also cooperate.
• Blood supply carries fuels between organs.
• Nervous and endocrine systems relay
messages between organs ensuring metabolic
pathways in individual organs are regulated to
meet needs of animal as a whole.
84
42
Case Study: Muscle Metabolism
REST vs. VIGOROUS EXERCISE
Slow release & use of Rapid mobilisation &

stored energy use of stored energy
How do we get this
rapid release of
energy?
85
Muscle at Rest
Major fuel is fatty acids (FA)
Triacylglycerol
(TAG)
lipase
FA are degraded to
acetyl CoA &
glycerol +
BLOOD metabolised via TCA
FA
(FA carried on cycle & OxPhos
serum albumin;
delivery is slow) Muscle
Adipose tissue
86
43
Resting Muscle has Plenty of O2
A process of sequential release
FA of 2‐C acetyl CoA units
β oxidation
[H]
Acetyl CoA ATP
OAA H2O
CAC
O2
[H] ETC
87
Muscle during Vigorous Exercise
• ATP consumption rapidly
increases (x 100)
• ATP resynthesis becomes
limiting
• FA oxidation is now
inadequate (i.e. insoluble
fats are slowly mobilised
and O2 supply to muscle
also becomes limiting)
Muscle glycogen is metabolised by anaerobic glycolysis
88
44
Metabolism in Active Muscle
Glycogen
Blood
Glucose
ANAEROBIC glycolysis
supplies ATP
Glucose‐6‐phosphate
2 ATP (or 3 ATP if
from glycogen)
Pyruvate Pyruvate shunted to lactate to
LDH regenerate NAD+
Lactate
Acetyl CoA TCA cannot process all acetyl
CoA due to insufficient O2
89
The Cori Cycle
Lactate is produced in muscle via anaerobic
glycolysis. Replenishes NAD+
Lactate is released into the blood stream and
is carried to the liver.
Lactate builds up during vigorous exercise
(increase in [lactate] in muscle & blood, &
movement to liver, occurs relatively slowly)
Time delay occurs during recycling of lactate
to glucose (need for O2 is delayed) "O2 debt”
In liver, it is reconverted to pyruvate and then
to glucose by the process of gluconeogensis.
Oxygen is required for this process.
90
45
Review of the Roles of Systems
in Muscle Contraction
Fig 25‐1: Energy metabolism in skeletal muscle
91
Feeding & Fasting: Maintaining
Blood [Glucose] is Critical
Blood [glucose] is maintained within very narrow
limits, ~ 4‐7 mM.
 Reason‐ many cells require constant supply of glucose
 If blood glucose drops below 2‐3 mM causing loss of
consciousness and even death.
 If blood [glucose] are > 10 mM glucose is lost in urine
causing dehydration & other complications.
Body maintains blood glucose concentration by:
 removing glucose from blood & storing after a meal.
 replacing blood glucose from body stores during fasting.
92
46
Insulin↑ Body cells take up
Glucagon ↓ more glucose.
pancreas
releases insulin
into the blood.
Liver takes
up glucose
and stores it
as glycogen.
Maintenance STIMULUS:
[glucose] increases
after carbohydrate‐
[GLUCOSE] DECLINES
stimulus for insulin
release diminishes
rich meal)
of Glucose Blood glucose level
Homeostasis
(5mM)
[GLUCOSE] RISES STIMULUS:
stimulus for glucagon [glucose] decreases
release diminishes. (after skipping meal)
Liver breaks
down glycogen
& releases pancreas
glucose into releases glucagon
blood. into the blood.
Insulin ↓
Glucagon ↑
93
Target Tissues for Insulin & Glucagon
Insulin reduces blood glucose levels by
 Promoting the cellular uptake of glucose.
 Slowing glycogen breakdown in the liver.
 Promoting fat storage in adipose tissue.
Glucagon increases blood glucose levels by
 Stimulating the conversion of glycogen to glucose in the
liver not muscle.
 Stimulating the breakdown of fat and the conversion of
protein (NOT fat) into glucose (gluconeogenesis).
Adrenaline also acts in the short‐term (fight or flight response)
94
47
Blood Glucose Concentrations
90
80
Unregulated
70
Blood Glucose (mM)
60
50
40
30
Regulated by insulin
20
10
0
0 2 4 6 8 10 12 14 16 18 20 22 24
Time (h)
Meal
95
The Fed State
Amounts of fuel molecules stored in a "typical" person
compact energy store; can last many
Fat ~15 kg
months; but not rapidly mobilized.
~25% in liver, ~75% in muscle, ~24 hrs
Glycogen 0.2 ‐ 0.5 Kg

reserve only, rapidly mobilised.
no specialised storage proteins in
animals (main “store” is muscle);
Protein 15 Kg muscles are "wasted" during long‐
term starvation; converted to glucose
(via gluconeogenesis) & KB.
NOTE: Some tissues (brain, red blood cells) must have glucose
for energy.
96
48
Metabolism in the Fed State
• After a meal, food is oxidised to provide energy.
• In addition, the hormone insulin promotes the storage of
food.
Carbohydrates Fat Protein
Glucose Fatty acids Amino acids
Storage Glycogen Fat Protein
CO2, H2O, energy
97
Hormonal Control of
Metabolism in the Fed State
Glucose
+
Glucose
transporter
Glucose
+
Glycogen (Storage)
+
Acetyl‐CoA
+
+ Activated/increased
CAC by insulin
Fat
(Energy)
(Storage)
98
49
Glycogen
Glycogen is a storage form of glucose in liver & muscle.
Cells store glucose as glycogen to decrease osmotic
pressure.
LIVER (not muscle) glycogen is a short‐term glucose
reserve. Helps maintain blood glucose during fasting.
Muscle glycogen is energy reserve during EXERCISE, not
fasting.
An increase in the concentration of insulin promotes the
synthesis of glycogen from glucose in liver and muscle.
99
Fed State ‐ Carbohydrate Metabolism
+ Promoted Liver
by insulin Fatty acid
TAG + Glycogen
+
Glucose
TAG Glucose +
Glucose + Blood Glucose
Fatty +
acid
CO2, H2O Glycogen
Adipose
tissue Muscle
Brain
100
50
Fasting State: The Liver ‐ 1
Glucose
Glucose
transporter + +
Glucose Glycogen
+
Pyruvate
+
+ Activated by
Amino acids glucagon
101
Fasting State: The Liver ‐ 2
Glucose
Glucose
transporter
Glucose
+
Glycogen (Storage)
X NO
Acetyl‐CoA
+ +
CAC Activated by
Fat
(Energy) glucagon
(Storage)
102
51
Metabolism in the Fasting State
Glycogen
Amino FA
acids
FA
Amino Ketone
acids bodies
Glucose TAG
Protein
CO2
CO2
103
Gluconeogenesis Maintains
Blood Glucose During Starvation
100
• Glycogen stores are
90
limited
80
• Gluconeogenesis
Percentage of blood glucose
70
becomes important for 60
supplying glucose after 50
about 24 h fasting 40
• Takes place in liver and 30
kidney 20
10
• Activated by glucagon
0
Inhibited by insulin 4 h 8 h 16 h 30 h
Glycogenolysis Gluconeogenesis
104
52
Substrates for Gluconeogenesis
• Glycerol from fat breakdown ‐ 25%
• Lactate from anaerobic glycolysis ‐ 25%
• Protein breakdown ‐ amino acids ‐ 50%
• Protein breakdown is triggered by fall in
[insulin] and rise in [glucagon].
• NOT fatty acids
105
Short‐term Fasting
Glycogen Protein
(Liver) (Muscle) Fat
(Adipose tissue)
Glucose Amino acids
Blood 5 C or 6 C Acetyl CoA

Glucose
Gluconeogenesis
CAC CAC
Oxaloacetate
CO2 CO2
ATP ATP
106
53
Acetyl CoA from Fat Cannot be
Converted to Glucose
 Oxaloacetate (from CAC cycle) is starting point for
glucose synthesis
 Acetyl CoA is burned totally to 2 CO2 during one
turn of CAC cycle
 Therefore acetyl CoA cannot be converted to
oxaloacetate
 Cannot reverse PDH
X
Acetyl CoA
OAA
CAC
2 CO2
107
Long‐term Fasting
FAT BODY PROTEIN
Lipase
Glycogen
Adipose
stores are Muscle
depleted
FA Amino acids
Liver  oxidation Liver
5 C or 6 C
Acetyl CoA
Oxaloacetate CAC
Ketone bodies
acetoacetate &
‐hydroxybutyrate
Glucose
Heart, brain, muscle Brain, RBC
108
54
• If you knew that times of nutritional deprivation
were coming, from an energy perspective, would
it be best to stock up on carbohydrate, fat or
protein?
• What are the reasons for your choice?
• We use lots of glucose, we also can make glucose.
How do we avoid futile cycling?
• Lactic acid is a substrate for gluconeogenesis –
does this route provide for a nett gain in glucose?
109
55

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BIOL1007 Notes - 2016

Lehninger 3rd Ed, p 487.

Food + O2 CO2 + H2O

Redox reactions are reactions Oxidation Involves Loss – Reduction Involves Gain

Donor‐e‐ + Acceptor ö Donor + Acceptor‐e‐

BIOL1007: Metabolism Module Glucose Oxidation ‐ Animation

Aerobic Fate of Pyruvate

Section 16.1 in Lehninger

Central Role of Acetyl CoA

The Tricarboxylic Acid Cycle

FADH2 + 0.5 O2 FAD + H2O via ETC

BUT ATP is only gained from NADH and FADH2 by

Slow release & use of Rapid mobilisation &

Glycogen 0.2 ‐ 0.5 Kg

Carbohydrates Fat Protein

Glucose Fatty acids Amino acids

Storage Glycogen Fat Protein

• Protein breakdown ‐ amino acids ‐ 50%

Blood 5 C or 6 C Acetyl CoA

Liver  oxidation Liver

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