Mitochondria respiratory electron-transport

The oxidation reactions involved are catalyzed by a series of nicotinamide adenine dinucleotide (NAD+) or flavin adenine dinucleotide (FAD) dependent dehydrogenases in the highly conserved metabolic pathways of glycolysis, fatty acid oxidation and the tricarboxylic acid cycle, the latter two of which are localized to the mitochondrion, as is the bulk of coupled ATP synthesis. Reoxidation of the reduced cofactors (NADH and FADH2) requires molecular oxygen and is carried out by protein complexes integral to the inner mitochondrial membrane, collectively known as the respiratory, electron transport, or cytochrome, chain. Ubiquinone (UQ), and the small soluble protein cytochrome c, act as carriers of electrons between the complexes (Fig. 13.1.1). 

Current estimates are that three protons move into the matrix through the ATP-synthase for each ATP that is synthesized. We see below that one additional proton enters the mitochondrion in connection with the uptake of ADP and Pi and export of ATP, giving a total of four protons per ATP. How does this stoichiometry relate to the P-to-O ratio When mitochondria respire and form ATP at a constant rate, protons must return to the matrix at a rate that just balances the proton efflux driven by the electron-transport reactions. Suppose that 10 protons are pumped out for each pair of electrons that traverse the respiratory chain from NADH to 02, and 4 protons move back in for each ATP molecule that is synthesized. Because the rates of proton efflux and influx must balance, 2.5 molecules of ATP (10/4) should be formed for each pair of electrons that go to 02. The P-to-O ratio thus is given by the ratio of the proton stoichiometries. If oxidation of succinate extrudes six protons per pair of electrons, the P-to-O ratio for this substrate is 6/4, or 1.5. These ratios agree with the measured P-to-O ratios for the two substrates. 

In eukaryotes, most of the reactions of aerobic energy metabolism occur in mitochondria. An inner membrane separates the mitochondrion into two spaces the internal matrix space and the intermembrane space. An electron-transport system in the inner membrane oxidizes NADH and succinate at the expense of 02, generating ATP in the process. The operation of the respiratory chain and its coupling to ATP synthesis can be summarized as follows ... 

Oxidative phosphorylation is ATP synthesis linked to the oxidation of NADH and FADH2 by electron transport through the respiratory chain. This occurs via a mechanism originally proposed as the chemiosmotic hypothesis. Energy liberated by electron transport is used to pump H+ ions out of the mitochondrion to create an electrochemical proton (H+) gradient. The protons flow back into the mitochondrion through the ATP synthase located in the inner mitochondrial membrane, and this drives ATP synthesis. Approximately three ATP molecules are synthesized per NADH oxidized and approximately two ATPs are synthesized per FADH2 oxidized. 

Mitochondrion—The site for the electron-transport and the respiratory enzyme systems in eucaryotes. 

The proteins of the respiratory chain are NADH dehydrogenase, cytochrome b, cytochrome Cj, cytochrome c, cytochrome aj, and cytochrome a. Cytochromes ai and as form a complex known as cytochrome c oxidase. The proteins are listed in the order in which they are used in the electron transport pathway. The proteins are all membrane-bound proteins, though cytochrome c is only weakly bound to the outer surface of the irmer membrane. Its polypeptide chain is not inserted in the membrane. Electrons are delivered in pairs, via NAD, to-the respiratory chain. It is thought that three protons are driven out of the mitochondrion for each pair of electrons from NAD passing down the respiratory chain and through cytochrome oxidase to oxygen (Cross, 1981 Hatefi, 1985). Further details are given in Chapter 5 and xmder Iron in Chapter 10. 

Maintenance of respiratory control depends on the structural integrity of the mitochondrion. Disruption of the organelle causes electron transport to become uncoupled from ATP synthesis. Under these conditions, oxygen uptake proceeds at high rates under all conditions. ATP synthesis is inhibited, even though electrons are being passed along the respiratory chain and used to reduce 02 to water. 

The degree of conservation, in terms of subunit composition and protein sequence, between mammalian respiratory chain complexes and those characterized from fungi and other organisms depends on the subunit and complex being considered (detailed in specific sections below), but in general, those subunits which are known to have a central role in electron transport are well conserved in terms of protein sequence and, where known, tertiary structure. For these subunits, a dear relationship to bacterial respiratory chain components can also be seen, which leads to the condusion that the mitochondrial respiratory chain complexes have evolved and adapted from those of the symbiotic bacterial ancestor of the mitochondrion [23]. Mitochondrial complexes have in most cases acquired many additional subunits whose function remains obscure. 

The inner membrane of the mitochondrion accounts for 80-95% of the protein found in mitochondrial membranes and over 90% of the lipid. It is the site of the respiratory chain and the synthesis of ATP. It is this membrane, in conjunction with studies on transport through the plasma membrane, that has contributed most forcefully both to the viewpoint of the anisotropic organization of membrane structural elements and of biochemical events carried out by or in membranes. As regards mitochondria, the interaction of the inner membrane components in carrying out electron transport and oxidative phosphorylation is the focal investigative question both for mitochondrial function and for the organization of vectorial events in general. 

Electron transport and oxidative phosphorylation represent the most complex membrane processes yet uncovered in living mammalian cells. The broad outlines of this pathway are not much in question. Thus, the oxidation of one molecule of NADH by the respiratory chain results in the formation of three molecules of ATP. Three complexes along this chain—NADH-Q reductase, QH2-cytochrome c reductase, and cytochrome c oxidase—contain the sites where energy is transduced and enabled to interact with the ATPase to generate ATP (Fig. 2). Abundant evidence exists, as well, for the transfer of energy among these three complexes without the involvement of ATP synthesis for instance in ion translocation by the mitochondrion (Emster, 1977). 

Mitchell based his concept on the suggestion that as electron is transported along the respiratory chain, H+ ions are ejected to cytoplasm (the mitochondrion environment). As a consequence, a gradient of H+ ion concentration occurs in external and internal mitochondrial spaces. Of course, this H+ ion concentration gradient is supported by electron transfer free energy decrease and in the case of membrane impermeability for H+ ions. 

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