Oxidative Phosphorylation in Eukaryotes Takes Place in Mitochondria

Mitochondria generate most of the ATP required by aerobic cells by a joint endeavor of the reactions of citric acid cycle, which take place in the mitochondrial matrix, and oxidative phosphorylation, which takes place in the inner mitochondrial membrane. Mitochondria are descendents of a free-living bacterium that established a symbiotic relation with another cell.

Oxidative Phosphorylation Depends on Electron Transfer

In oxidative phosphorylation, the synthesis of ATP is coupled to the flow of electrons from NADH or FADH₂ to O₂ by a proton gradient across the inner mitochondrial membrane. Electron flow through three asymmetrically oriented transmembrane complexes results in the pumping of protons out of the mitochondrial matrix and the generation of a membrane potential. ATP is synthesized when protons flow back to the matrix through a channel in an ATP-synthesizing complex, called ATP synthase (also known as F₀F₁-ATPase). Oxidative phosphorylation exemplifies a fundamental theme of bioenergetics: the transmission of free energy by proton gradients.

The Respiratory Chain Consists of Four Complexes: Three Proton Pumps and a Physical Link to the Citric Acid Cycle

The electron carriers in the respiratory assembly of the inner mitochondrial membrane are quinones, flavins, iron-sulfur complexes, heme groups of cytochromes, and copper ions. Electrons from NADH are transferred to the FMN prosthetic group of NADH-Q oxidoreductase (Complex I), the first of four complexes. This oxidoreductase also contains Fe-S centers. The electrons emerge in QH₂, the reduced form of ubiquinone (Q). The citric acid cycle enzyme succinate dehydrogenase is a component of the succinate-Q reductase complex (Complex II), which donates electrons from FADH₂ to Q to form QH₂.This highly mobile hydrophobic carrier transfers its electrons to Q-cytochrome c oxidoreductase (Complex III), a complex that contains cytochromes b and c₁ and an Fe-S center. This complex reduces cytochrome c, a water-soluble peripheral membrane protein. Cytochrome c, like Q, is a mobile carrier of electrons, which it then transfers to cytochrome c oxidase (Complex IV). This complex contains cytochromes a and a₃ and three copper ions. A heme iron ion and a copper ion in this oxidase transfer electrons to O₂, the ultimate acceptor, to form H₂O.

A Proton Gradient Powers the Synthesis of ATP

The flow of electrons through Complexes I, III, and IV leads to the transfer of protons from the matrix side to the cytosolic side of the inner mitochondrial membrane. A proton-motive force consisting of a pH gradient (matrix side basic) and a membrane potential (matrix side negative) is generated. The flow of protons back to the matrix side through ATP synthase drives ATP synthesis. The enzyme complex is a molecular motor made of two operational units: a rotating component and a stationary component. The rotation of the γ subunit induces structural changes in the β subunit that result in the synthesis and release of ATP from the enzyme. Proton influx provides the force for the rotation.

The flow of two electrons through NADH-Q oxidoreductase, Q-cytochrome c oxidoreductase, and cytochrome c oxidase generates a gradient sufficient to synthesize 1, 0.5, and 1 molecule of ATP, respectively. Hence, 2.5 molecules of ATP are formed per molecule of NADH oxidized in the mitochondrial matrix, whereas only 1.5 molecules of ATP are made per molecule of FADH₂ oxidized because its electrons enter the chain at QH₂, after the first proton-pumping site.

Many Shuttles Allow Movement Across the Mitochondrial Membranes

Mitochondria employ a host of carriers, or transporters, to move molecules across the inner mitochondrial membrane. The electrons of cytoplasmic NADH are transferred into the mitochondria by the glycerol phosphate shuttle to form FADH₂ from FAD. The entry of ADP into the mitochondrial matrix is coupled to the exit of ATP by ATP-ADP translocase, a transporter driven by membrane potential.

The Regulation of Oxidative Phosphorylation Is Governed Primarily by the Need for ATP

About 30 molecules of ATP are generated when a molecule of glucose is completely oxidized to CO₂ and H₂O. Electron transport is normally tightly coupled to phosphorylation. NADH and FADH₂ are oxidized only if ADP is simultaneously phosphorylated to ATP, a form of regulation called acceptor or respiratory control. Uncouplers such as DNP can disrupt this coupling; they dissipate the proton gradient by carrying protons across the inner mitochondrial membrane. Proteins have been identified that uncouple electron transport and ATP synthesis for the generation of heat.

Key Terms

oxidative phosphorylation

proton-motive force

cellular respiration

electron-transport chain

reduction (redox, oxidation-reduction, E´₀) potential

inverted region

coenzyme Q (Q, ubiquinone)

NADH-Q oxidoreductase (Complex I)

flavin mononucleotide (FMN)

iron-sulfur (nonheme iron) protein

succinate-Q reductase (Complex II)

Q-cytochrome c oxidoreductase (Complex III)

cytochrome c (cyt c)

Rieske center

Q cycle

cytochrome c oxidase (Complex IV)

superoxide dismutase

catalase

ATP synthase (Complex V, F₁F₀ ATPase)

glycerol 3-phosphate shuttle

malate-aspartate shuttle

ATP-ADP translocase (adenine nucleotide translocase, ANT)

respiratory (acceptor) control

uncoupling protein (UCP)

programmed cell death (apoptosis)

caspase