20-4 The Connection between Electron Transport and Phosphorylation

 Some of the energy released by the oxidation reactions in the electron transport chain is used to
drive the phosphorylation of ADP.
 The phosphorylation of each mole of ADP requires 30.5 kJ = 7.3 kcal, and we have seen how
each reaction catalyzed by three of the four respiratory complexes provides more than enough
energy to drive this reaction, although it is by no means a direct usage of this energy.
 The energy of the electrochemical potential (voltage drop) across the membrane is converted to
the chemical energy of ATP by the coupling process.

 What is the coupling factor in oxidative phosphorylation?

 A complex protein oligomer, separate from the electron transport complexes, serves this
function; the complete protein spans the inner mitochondrial membrane and projects into
the matrix as well. The portion of the protein that spans the membrane is called F 0.
 The portion that projects into the matrix is called F 1; it consists of five different kinds of
polypeptide chains.
 ATP Synthase- the enzyme responsible for production of ATP in mitochondria.

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 Uncouplers- substances that overcome the proton gradient in mitochondria, allowing

electron transport to proceed in the absence of phosphorylation.
 A well-known example of an uncoupler is 2,4-dinitrophenol. Various antibiotics, such
as valinomycin and gramicidin A, are also uncouplers.
  P / O Ratio- the ratio of ATP produced by oxidative phosphorylation to oxygen atoms
consumed in electron transport.

20-5 The Mechanism of Coupling in Oxidative Phosphorylation

 Several mechanisms have been proposed to account for the coupling of electron transport
and ATP production. The mechanism that served as the point of departure in all discussions
is chemiosmotic coupling, which was later modified to include a consideration of
conformational coupling.

Chemiosmotic Coupling Mechanism

 What Is chemiosmotic coupling?
 Chemiosmotic Coupling- the mechanism for coupling electron transport to oxidative
phosphorylation, it requires a proton gradient across the inner mitochondrial membrane.
 The proton gradient exists because the various proteins that serve as electron carriers in the
respiratory chain are not symmetrically oriented with respect to the two sides of the inner
mitochondrial membrane, nor do they react in the same way with respect to the matrix and
the intermembrane space.

 Dinitrophenol is an acid; its conjugate base, dinitrophenolate anion, is the actual uncoupler
because it can react with pro-tons in the intermembrane space, reducing the difference in
proton concentration between the two sides of the inner mitochondrial membrane.
 Ionophores- substances that create channels for ions to pass through the inner
mitochondrial membrane.

Conformational Coupling Mechanism

 Conformational Coupling- a mechanism for coupling electron transport to oxidative
phosphorylation that depends on a conformational change in the ATP synthetase.
 Three sites for substrate on the synthase and three possible conformational states:
1. open (O), with low affinity for substrate
2. loose-binding (L), which is not catalytically active
3. tight-binding (T), which is catalytically active

 A proton flux converts the site in the T conformation to the O conformation, releasing the
ATP.
 Electron micrographs have shown that the conformation of the inner mitochondrial
membrane and cristae is distinctly different in the resting and active states. It is well
established that the shape of mitochondria is not static.