Electron transport side reactions

The second electron shuttle system, called the malate-aspartate shuttle, is shown in Figure 21.34. Oxaloacetate is reduced in the cytosol, acquiring the electrons of NADH (which is oxidized to NAD ). Malate is transported across the inner membrane, where it is reoxidized by malate dehydrogenase, converting NAD to NADH in the matrix. This mitochondrial NADH readily enters the electron transport chain. The oxaloacetate produced in this reaction cannot cross the inner membrane and must be transaminated to form aspartate, which can be transported across the membrane to the cytosolic side. Transamination in the cytosol recycles aspartate back to oxaloacetate. In contrast to the glycerol phosphate shuttle, the malate-aspartate cycle is reversible, and it operates as shown in Figure 21.34 only if the NADH/NAD ratio in the cytosol is higher than the ratio in the matrix. Because this shuttle produces NADH in the matrix, the full 2.5 ATPs per NADH are recovered. 

Much of the difficulty in demonstrating the mechanism of breakaway in a particular case arises from the thinness of the reaction zone and its location at the metal-oxide interface. Workers must consider (a) whether the oxide is cracked or merely recrystallised (b) whether the oxide now results from direct molecular reaction, or whether a barrier layer remains (c) whether the inception of a side reaction (e.g. 2CO - COj + C)" caused failure or (d) whether a new transport process, chemical transport or volatilisation, has become possible. In developing these mechanisms both arguments and experimental technique require considerable sophistication. As a few examples one may cite the use of density and specific surface-area measurements as routine of porosimetry by a variety of methods of optical microscopy, electron microscopy and X-ray diffraction at reaction temperature of tracer, electric field and stress measurements. Excellent metallographic sectioning is taken for granted in this field of research. 

P. Mitchell (Nobel Prize for Chemistry, 1978) explained these facts by his chemiosmotic theory. This theory is based on the ordering of successive oxidation processes into reaction sequences called loops. Each loop consists of two basic processes, one of which is oriented in the direction away from the matrix surface of the internal membrane into the intracristal space and connected with the transfer of electrons together with protons. The second process is oriented in the opposite direction and is connected with the transfer of electrons alone. Figure 6.27 depicts the first Mitchell loop, whose first step involves reduction of NAD+ (the oxidized form of nicotinamide adenosine dinucleotide) by the carbonaceous substrate, SH2. In this process, two electrons and two protons are transferred from the matrix space. The protons are accumulated in the intracristal space, while electrons are transferred in the opposite direction by the reduction of the oxidized form of the Fe-S protein. This reduces a further component of the electron transport chain on the matrix side of the membrane and the process is repeated. The final process is the reduction of molecular oxygen with the reduced form of cytochrome oxidase. It would appear that this reaction sequence includes not only loops but also a proton pump, i.e. an enzymatic system that can employ the energy of the redox step in the electron transfer chain for translocation of protons from the matrix space into the intracristal space. 

First isolated from human urine, biopterin (Fig. 15-17) is present in liver and other tissues where it functions in a reduced form as a hydroxylation coenzyme (see Chapter 18).338 It is also present in nitric oxide synthase (Chapter 18).341/342 Other functions in oxidative reactions, in regulation of electron transport, and in photosynthesis have been proposed.343 Neopterin, found in honeybee larvae, resembles biopterin but has a D-erythro configuration in the side chain. The red eye pigments of Drosophila, called drosopterins, are complex dimeric pterins containing fused 7-membered rings (Fig. 15-17).344 345... 

The TCA cycle is carefully regulated to ensure that its level of activity relates to cellular needs. The cycle serves two functions (1) furnishing reducing equivalents (as NADH and to a lesser extent as FADH2) to the electron-transport chain, and (2) by means of side reactions, providing substrates for biosynthesis reactions. Both of these functions are reflected in the regulation of the cycle. 

Fig. 5.9. Proposed scheme for the intramitochondrial metabolism of malate by Hymenolepis diminuta. Abbreviations ME, malic enzyme F, fumarase T, transhydrogenase FR, fumarate reductase ETS, electron transport system. Once within the matrix compartment, malate oxidation, as catalysed by malic enzyme, results in NADPH formation. Via the activity of fumarase, malate also is converted to fumarate in the matrix compartment. NADPH then serves as a substrate for the inner-membrane-associated transhydrogenase and transhydrogenation between NADPH and matrix NAD is a scalar reaction associated with the matrix side of the inner membrane. Matrix NADH so formed reduces the electron transport system via a site on the matrix side of the inner membrane permitting fumarate reductase activity. The reduction of fumarate to succinate results in succinate accumulation within the matrix compartment. (After McKelvey Fioravanti, 1985.)...

Apart from the production of NADH and FADH2, which are the high-energy fuels of electron transport, the citric acid cycle has two other major functions. Several of its intermediate compounds are used to synthesize other cell constituents. This, the provision of molecules for other metabolic or biosynthetic pathways, is the anabolic function of the cycle (Table 12.1). Alternatively, certain other processes occurring within the cell may produce intermediates of the citric acid cycle. These compounds enter the reactions of the cycle, and their degradation involves the catabolic role of the cycle. These two major capabilities classify the citric acid cycle as an amphibolic pathway (Greek amphi meaning both sides ). 

Figure 18.21. Proton Transport by Cytochrome C Oxidase. Four "chemical" protons are taken up from the matrix side to reduce one molecule of O2 to two molecules of H2O. Four additional "pumped" protons are transported out of the matrix and released on the cytosolic side in the course of the reaction. The pumped protons double the efficiency of free-energy storage in the form of a proton gradient for this final step in the electron-transport chain.

The dilemma posed by these considerations was resolved for the DHP-organized system when it was noted that upon illumination the initially compartmented MV + partially leaked out of the vesicles [108]. Thus, the observed reaction actually occurred between reactants adsorbed on the same vesicle surface. The mechanism of this unprecedented light-induced scrambling of MV + is still not understood. Likewise, subsequent investigations of the PC-organized system provided evidence for transmembrane leakage of a small amount of the compartmented (C7)2V + ion, which then facilitated transmembrane electron transport [109]. Specifically, the reaction characteristics were duplicated when the amphiphilic Ru(bpy)3 + analog was bound only to the opposite side of the membrane as the oxidative quencher... 

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