Topic 10-1 – The Citric Acid Cycle

Text book – Chapter 14

Essential Biochemistry
Third Edition
Charlotte W. Pratt | Kathleen Cornely
Online: Interactive Concepts in Biochemistry
http://vcell.ndsu.nodak.edu/animations/citricacid_overview/first.htm
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 - 2:15

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 The Pyruvate Dehydrogenase
Reaction
http://www.wiley.com/college/boyer/0470003790/animations/pdc/pdc.htm

Surface View Cutaway View

Model of the pyruvate

dehydrogenase complex
from
B. stearothermophilus

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 The Pyruvate Dehydrogenase
Reaction
http://www.wiley.com/college/boyer/0470003790/animations/pdc/pdc.htm

Where does this occur? Mitochondrial matrix

How does it get into the Mitochondria? Active transport via pyruvate translocase

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 KEY CONCEPTS: Section 14-1
• The pyruvate dehydrogenase complex
includes three types of enzymes that
collectively remove a carboxylate group
from pyruvate and produce acetyl-CoA
and NADH.

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Overview of
the Citric
Acid Cycle
• The CoA released can be
reused by the pyruvate
dehydrogenase complex or
used later in the citric acid
cycle to synthesize the
intermediate succinyl-CoA.
• 1 glucose → 2 Acetyl-CoA
→ 2 revolutions for each
glucose

acetyl-CoA + GDP + Pi + 3 NAD+ + Q 2 CO2 + CoA + GTP + 3 NADH + QH2

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 Fate of
Carbons in
the Citric
Acid Cycle
Carbon
• The two carbons Carbon from
Acetyl-CoA
released in the 1st
round of the citric
acid cycle did not
come from acetyl-
CoA.

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 The citric acid
cycle is an
energy-
generating cycle.

• 1 NADH = 2.5 ATP

• 1 QH2 = 1.5 ATP

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 Regulation occurs at the three
irreversible steps.
• The product of the 1st reaction, citrate, inhibits
citrate synthase

• Succinyl-CoA, the product of Reaction 4, inhibits

the enzyme that produces it. It also acts as a
feedback inhibitor by competing with acetyl-CoA in
Reaction 1.

• The activity of isocitrate dehydrogenase

(Reaction 3) is inhibited by its reaction product,
NADH. NADH also inhibits a-ketoglutarate
dehydrogenase and citrate synthase.

• Both dehydrogenases are activated by Ca-ions,

which generally signify the need to generate
cellular free energy. ADP, also representing the
need for more ATP, activates isocitrate
dehydrogenase.

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 KEY CONCEPTS: Section 14-2

• The citric acid cycle is a set of eight reactions in

which an acetyl group is condensed with
oxaloacetate, two CO2 are lost, and
oxaloacetate is regenerated.
• Each round of the citric acid cycle generates
three NADH, one QH2, and one GTP or ATP.
• Flux through the citric acid cycle is regulated
primarily by feedback inhibition at three steps.

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 ANABOLIC AND CATABOLIC FUNCTIONS:
intermediates are precursors of other molecules.

Thus the cycle is amphibolic, serving both catabolism and anabolism.

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 An Example of Amino Acid
Formation from a Citric Acid Cycle
Intermediate

Arginine
Proline
Glutamine
Nucleotides
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 Intermediates that are diverted to
other pathways can be replenished.

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Anaplerotic reactions replenish
citric acid cycle intermediates.

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During exercise, some pyruvate can
be converted to α-ketoglutarate to
boost the activity of the citric acid
cycle via pyruvate carboxylase.

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 Citrate
Transport
System
• Citrate and
pyruvate
can cross the
mitochondrial
membrane via
specific transport
proteins.

• Makes acetyl-CoA
available for fatty
acid synthesis

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 KEY CONCEPTS: Section 14-3
• The citric acid cycle also supplies
precursors for the synthesis of other
compounds – it is thus an amphibolic
pathway.

• Citric acid cycle intermediates can be

replenished by anaplerotic reactions.

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 2:15 -

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Topic 10-2 – Electron Transfer Chain
and Oxidative Phosphorylation
Text book – Chapter 15

Essential Biochemistry
Third Edition
Charlotte W. Pratt | Kathleen Cornely
Online: Interactive Concepts in Biochemistry
http://vcell.ndsu.nodak.edu/animations/etc/index.htm
http://vcell.ndsu.nodak.edu/animations/atpgradient/index.htm

© 2014 John Wiley & Sons, Inc. All rights reserved.

 Overview in
Context

Oxidative phosphorylation is the

metabolic pathway in which cells
use enzymes to oxidize nutrients,
thereby releasing energy which is
used to reform ATP. In most
eukaryotes, this takes place inside
mitochondria.

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 Recap on Oxidation-Reduction

• One reactant is in its oxidized state while

the other is in its reduced state.
• Loss of electrons = oxidation
• Gain of electrons = reduction
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Reduction potential
indicates a substance’s
tendency to accept
electrons.

Large positive values –

strong oxidizing agents
– easily accepts
electrons

Large negative values –

strong reducing agents
– easily donates
electrons

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 KEY CONCEPTS: Section 15-1
• The standard reduction potential indicates a
substance’s tendency to become reduced; the
actual reduction potential depends on the
concentrations of reactants.
• Electrons are transferred from a substance with
a lower reduction potential to a substance with
a higher reduction potential.
• The free energy change for an oxidation-
reduction reaction depends on the change in
reduction potential.

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 In eukaryotes electron transport
takes place in the mitochondrion.

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 The electron
transport chain
involves four
protein
complexes

O2

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 Overview of Mitochondrial Electron
Transport • 1 NADH = 2.5 ATP
• 1 QH2 = 1.5 ATP

Fig. 17.13 from Concepts in Biochemistry (Boyer)

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Complex I

• Catalyzes the transfer of a pair of

electrons from NADH to ubiquinone (Q),

• Transfers four protons from the matrix to

the intermembrane space

• The reduced quinone product of the

Complex I reaction joins a pool of
quinones that are soluble in the inner
mitochondrial membrane.

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Complex II

of the mitochondrial
respiratory chain is more like a tributary
because it does not undertake proton
translocation and therefore does not
directly contribute the free energy of its
oxidation–reduction reaction toward ATP
synthesis. Nevertheless, it does feed
reducing equivalents as ubiquinol into the
electron transport chain

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Complex III

• Ubiquinol is reoxidized

• Transfers electrons to the peripheral

membrane protein cytochrome c

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Complex IV

• is the last enzyme to deal with the electrons

derived from the oxidation of metabolic
fuels. Four electrons delivered by
cytochrome c are consumed in the reduction
of molecular oxygen to water

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 Video: 2:30

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 KEY CONCEPTS: Section 15-2
• The inner mitochondrial membrane encloses the
matrix and includes specific transport proteins.
• Complex I transfers electrons from NADH to
ubiquinone.
• The citric acid cycle, fatty acid oxidation, and
other processes also generate mitochondrial
ubiquinol.
• The Q cycle mediated by Complex III reduces
cytochrome c.
• Complex IV uses electrons from cytochrome c to
reduce O2 to H2O.
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 How much energy is available from
electron transport?
• 1 NADH = 2.5 ATP
• 1 QH2 = 1.5 ATP

• This is enough energy to drive the

endergonic phosphorylation of ADP to
form ATP (ΔG°' = +30.5 kJ•mol–1)!

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The imbalance of protons represents a
source of free energy, also called a
protonmotive force, that can drive the
activity of an ATP synthase.

[H+] = low

[H+] = high

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 KEY CONCEPTS: Section 15-3
• The formation of a transmembrane proton
gradient during electron transport
provides the free energy to synthesize
ATP.

• Both concentration and charge contribute

to the free energy of the proton gradient.

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 The protein that taps the
electrochemical proton gradient to
phosphorylate ADP is known as ATP
synthase (Complex V).

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 ATP Synthase Function
• Use the
electrochemical Form ATP
gradient to drive
phosphorylation.

• Uses mechanical energy

of a rotational
conformation change to
form a chemical bond
(endergonic reaction
ΔG°' = +30.5 kJ•mol–1)

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 KEY CONCEPTS: Section 15-4
• Proton translocation drives the rotation of a portion
of ATP synthase.
• Rotation-induced conformational changes allow
ATP synthase to bind ADP and Pi to phosphorylate
ADP, and to release ATP.
• The supply of reduced cofactors determines the
rate of oxidative phosphorylation.

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