Oxidative Phosphorylation

- Glycolysis + TCA cycle produces very little ATP directly (substrate level):
Glycolysis 2 ATPs and TCA cycle 2 ATPs per glucose
- The rest of the energy is stored in the reduced cofactors:
10 NADH + 2 FADH2 per glucose
- Consider that: C6H12O6 + 6 O2 6 CO2 + 6 H2O

(G' = -2870 kcal/mol)

- Oxidation of the reduced co-factors releases energy:

NADH + H+ + 1/2 O2 NAD+ + H2O (G' = -220 kcal/mol)
FADH2 + 1/2 O2 FAD + H2O

(G' = -182 kcal/mol)

- Thus, the reduced co-factors from oxidation of 1 mole of glucose store

10 X 220 + 2 X 182 = 2564 kJ of energy, accounting for about 90% of the
total realizable energy (under standard conditions)

NAD+/NADH and NADP+/NADPH

- Diffusible 2 electron carriers

- Accept or donate 1 hydride (H-) ion (1 proton and 2 electrons)

FAD/FADH2 and FMN/FMNH2

- Bound in enzymes as prosthetic groups

- 2 electron-carriers
- May transfer 1H+ + 1e- at a time

The mitochondrion

- Aerobic oxidation of biomolecules

Alligator jaw muscle - white muscle

Flight muscle - red muscle

Electron transport chain (ETC) in mitochondria

Electron transfer reactions
- The driving force is expressed as a difference in the standard redox
potentials of the components, and this is equivalent to the change in free
energy:
n = no. of e- transferred
G = -nF E'
F = 96.4 kJ mol-1 V-1
E = EA' - ED'

- Electron flow is favorable from donors of low potential (i.e. more negative
E values) to acceptors of high potential.
Four integral membrane protein complexes:

Complex I

Complex II

Complex III

Complex IV

NADH dehydrogenase (Complex I)

Cofactors (electron carriers): riboflavin coenzyme FMN, iron-sulfur clusters,
transfers electrons from NADH to unbound ubiquinone (Q)
the unbound ubiquinol (QH2) carries electrons diffuses through inner
membrane to cytochrome bc1 complex (Complex III)
Overall: NADH + H+ + Q NAD+ + QH2
(intermembrane
space)
Proton
pumping

(matrix)

FMN
- Protein-bound co-factor
- Accepts/donates 1 hydride ion
(1 electron + 1 proton) at a time
- 2 electron-carrier

Iron-sulfur (Fe-S) clusters

Single electron carriers: Fe2+ Fe3+ + e2Fe-2S:

Protein

4Fe-4S:

Protein

only the inorganic S is counted in the designation

each Fe is always coordinated by 4 S
4 cysteine residues from the protein also contribute 4S to the cluster
it is also posible to have a single Fe coordinated by 4 cysteine residues

Q (or CoQ)/QH2
- Diffusible through inner
membrane
- Accepts/donates 1 electron +
1 proton at a time
- A carrier of both electrons
(2) and protons (2)

Succinate dehydrogenase (Complex II)

The only membrane-bound enzyme in TCA cycle

Contains an internal chain of electron transfer cofactors
No proton pumping in Complex II

(intermembrane
space)

(matrix)

Cytochrome bc1 complex (Complex III)

Electron carriers: Fe-S clusters, cytochrome b and cytochrome c1

Transfers electrons from ubiquinone to cytochrome c
QH2 + 2 cyt c1 (oxidized) + 2 H+ Q + 2 cyt c1 (reduced) + 4H+
Cytochrome c is a water-soluble protein coenzyme in the
intermembrane space.
Complex III pumps 4 protons per 2 electrons transferred

(Iron-sulfur protein)
(intermembrane space)

(matrix)

Cytochromes

- Protein coenzymes
- Heme as a tightly bound co-factor
- Heme is a tetrapyrrole coordinating a single
atom of Fe(II/III)
- Single electron-carrier (Fe2+ Fe3+ + e-)

Cytochrome a

- Contains heme A
- Membrane-bound

Cytochrome b

- Contains heme B
- Membrane-bound

Cytochrome c

- Contains heme C
- Membrane-bound or
diffusible

The Q cycle in Complex III

Cytochrome oxidase (Complex IV)

Electron carriers: 2 Cu ions and 2 heme A groups (cytochrome a proteins)

Transfers electrons from cytochrome c to oxygen
4 cytochrome c (reduced) + 8H+ in + O2
4 cytochrome c (oxidized) + 4H+ out + 2 H2O
The redox centers only transfer electron one at a time
Incompletely reduced intermediates (e.g. hydrogen peroxide and hydroxyl
free radicals) remain tightly bound until complete reduction to water.
(x2)

(intermembrane
space)

(matrix)

Electron transport and proton pumps

inner
membrane

Proton gradient

(intermembrane
space)

(matrix)

A protein gradient is then established across the inner membrane

The energy stored in the gradient, proton-motive force, has two components:
chemical potential energy and electrical potential energy
The electrochemical energy released when protons flow spontaneously down
the gradient can be used to drive the synthesis of ATP from ADP and Pi

ATP synthesis by ATP synthase

ATP synthase: F0F1 complex in the inner membrane

Protons flow through the F0 unit down the gradient
ATP is synthesized by the F1 unit (ATPase) from ADP and Pi
For every 2 electrons donated by NADH, 2.5 ATPs are synthesized.
For every 2 electrons donated by FADH2, 1.5 ATPs are synthesized.

NADH + H+

NAD+

Shuttle systems for NADH generated in cytoplasm

Inner mitochondrial membrane is impermeable to NADH

Cytosolic NADH is shuttled indirectly into the mitochondria as reducing
equivalents
Reducing equivalents are molecules that can be transported into the
mitochondria

(1) Glycerol 3-P shuttle

- Skeletal muscle and brain
(DHAP)

Complex III
Complex IV

(2) Malate-aspartate shuttle

- Liver, kidney, and heart

(Complexes I, III, IV)

Net profit of aerobic metabolism

If we start from glucose:
- Glycolysis to 2 pyruvate yields:
Cytosol
2 ATP
2 X 2.5 (malate-aspartate shuttle) = 5 ATP
2 NADH
or 2 X 1.5 (glycerol 3-P shuttle) = 3 ATP

- Conversion of 2 pyruvate to 2 acetyl-CoA:

2NADH
= 2 X 2.5 = 5 ATP

Mitochondria

- Oxidation of 2 acetyl-CoA in the TCA cycle:

20 ATP
32 or 30
Net yield = __________
ATP for complete oxidation of glucose to CO2