Succinate transport

Kimmich, G.A., Randles, J., Bennett, E. (1991). Sodium dependent succinate transport by isolated chick intestinal cells. Am. J. Physiol. 260, Cl 151—Cl 157. 

How can the FADH2 generated by the succinate-Q-reductase complex participate in electron transport if it is not free to diffuse from the enzyme complex Does the oxidation of succinate transport protons ... 

Succinic acid and succinic anhydride are sold in 25-kg net polyethylene (PE) bags having cardboard box protection for the anhydride, in 70-liter (50-kg net) fiber dmms, and in 55-gaHon (275-lb 125-kg net) dmms. The two products must be stored in a fresh, dry, ventilated area. Succinic anhydride must be carefully protected from moisture during transportation and storage to avoid hydrolysis to succinic acid. 

The TCA cycle can now be completed by converting succinate to oxaloacetate. This latter process represents a net oxidation. The TCA cycle breaks it down into (consecutively) an oxidation step, a hydration reaction, and a second oxidation step. The oxidation steps are accompanied by the reduction of an [FAD] and an NAD. The reduced coenzymes, [FADHg] and NADH, subsequently provide reducing power in the electron transport chain. (We see in Chapter 24 that virtually the same chemical strategy is used in /3-oxidation of fatty acids.)... 

This is a crucial point because (as we will see) proton transport is coupled with ATP synthesis. Oxidation of one FADHg in the electron transport chain results in synthesis of approximately two molecules of ATP, compared with the approximately three ATPs produced by the oxidation of one NADH. Other enzymes can also supply electrons to UQ, including mitochondrial 5w-glyc-erophosphate dehydrogenase, an inner membrane-bound shuttle enzyme, and the fatty acyl-CoA dehydrogenases, three soluble matrix enzymes involved in fatty acid oxidation (Figure 21.7 also see Chapter 24). The path of electrons from succinate to UQ is shown in Figure 21.8. 

FIGURE 21.8 A probable scheme for electron flow in Complex II. Oxidation of succinate occurs with rednction of [FAD]. Electrons are then passed to Fe-S centers and then to coenzyme Q (UQ). Proton transport does not occur in this complex. 

Mitchell s chemiosmotic hypothesis. The ratio of protons transported per pair of electrons passed through the chain—the so-called HV2 e ratio—has been an object of great interest for many years. Nevertheless, the ratio has remained extremely difficult to determine. The consensus estimate for the electron transport pathway from succinate to Og is 6 H /2 e. The ratio for Complex I by itself remains uncertain, but recent best estimates place it as high as 4 H /2 e. On the basis of this value, the stoichiometry of transport for the pathway from NADH to O2 is 10 H /2 e. Although this is the value assumed in Figure 21.21, it is important to realize that this represents a consensus drawn from many experiments. 

Engelhardt s experiments in 1930 led to the notion that ATP is synthesized as the result of electron transport, and, by 1940, Severo Ochoa had carried out a measurement of the P/O ratio, the number of molecules of ATP generated per atom of oxygen consumed in the electron transport chain. Because two electrons are transferred down the chain per oxygen atom reduced, the P/O ratio also reflects the ratio of ATPs synthesized per pair of electrons consumed. After many tedious and careful measurements, scientists decided that the P/O ratio was 3 for NADH oxidation and 2 for succinate (that is, [FADHg]) oxidation. Electron flow and ATP synthesis are very tightly coupled in the sense that, in normal mitochondria, neither occurs without the other. 

Consider the oxidation of succinate by molecular oxygen as carried out via the electron transport pathway... 

Succinyl-CoA derived from propionyl-CoA can enter the TCA cycle. Oxidation of succinate to oxaloacetate provides a substrate for glucose synthesis. Thus, although the acetate units produced in /3-oxidation cannot be utilized in glu-coneogenesis by animals, the occasional propionate produced from oxidation of odd-carbon fatty acids can be used for sugar synthesis. Alternatively, succinate introduced to the TCA cycle from odd-carbon fatty acid oxidation may be oxidized to COg. However, all of the 4-carbon intermediates in the TCA cycle are regenerated in the cycle and thus should be viewed as catalytic species. Net consumption of succinyl-CoA thus does not occur directly in the TCA cycle. Rather, the succinyl-CoA generated from /3-oxidation of odd-carbon fatty acids must be converted to pyruvate and then to acetyl-CoA (which is completely oxidized in the TCA cycle). To follow this latter route, succinyl-CoA entering the TCA cycle must be first converted to malate in the usual way, and then transported from the mitochondrial matrix to the cytosol, where it is oxida-... 

This complex consists of four subunits, all of which are encoded on nuclear DNA, synthesized on cytosolic ribosomes, and transported into mitochondria. The succinate dehydrogenase (SDH) component of the complex oxidizes succinate to fumarate with transfer of electrons via its prosthetic group, FAD, to ubiquinone. It is unique in that it participates both in the respiratory chain and in the tricarboxylic acid (TC A) cycle. Defects of complex II are rare and only about 10 cases have been reported to date. Clinical syndromes include myopathy, but the major presenting features are often encephalopathy, with seizures and psychomotor retardation. Succinate oxidation is severely impaired (Figure 11). 

Flutolanil is an inhibitor of succinate dehydrogenase complex (Complex II), in the mitochondrial respiratory electron transport chain. ... 

In addition to these more-or-less well characterized proteins, iron is known to be bound to certain flavoproteins such as succinic dehydrogenase (20), aldehyde oxidase (27), xanthine oxidase (22) and dihydrooro-tate dehydrogenase (23). Iron is present and functional in non-heme segments of the electron transport chain but again no real structural information is at hand (24). 

Different organic acids, primarily lactic acid, have been successfully used for decontamination of whole livestock carcasses, and the application of different organic acids used for decontamination has also been tested in the fruit and vegetable industry. Organic acids other than lactic acid that are known to have bactericidal effects are acetic, benzoic, citric, malic, propanoic, sorbic, succinic and tartaric acids (Betts and Everis 2005). The antimicrobial action is due to a reduction in the pH in the bacterial environment, disruption of membrane transport, anion accumulation or a reduction in the internal pH in the cell (Busta et al., 2001). Many fruits contain naturally occurring organic acids. Nevertheless, some strains, for example E. coli 0157, are adapted to an acidic environment. Its survival, in combination with its low infective dose, makes it a health hazard for humans. 

In the presence of the inhibitor rotenone (to prevent the oxidation of NADH by the electron transport chain), succinate can be metabolized only to fumarate, producing an FADH2 in the process. 

In the absence of rotenone, the NADH that is made from the conversion of succinate to oxaloacetate can be oxidized by the electron transport chain. The metabolism of succinate then becomes... 

Rotenone inhibits the transfer of electrons from NADH into the electron transport chain. The oxidation of substrates that generate NADH is, therefore, blocked. However, substrates that are oxidized to generate FADH2 (such as succinate or a-glycerol phosphate) can still be oxidized and still generate ATP. Because NADH oxidation is blocked, the NADH pool becomes more reduced in the presence of rotenone since there s nowhere to transfer the electrons. 

Ubiquinones (coenzymes Q) Q9 and Qi0 are essential cofactors (electron carriers) in the mitochondrial electron transport chain. They play a key role shuttling electrons from NADH and succinate dehydrogenases to the cytochrome b-c1 complex in the inner mitochondrial membrane. Ubiquinones are lipid-soluble compounds containing a redox active quinoid ring and a tail of 50 (Qio) or 45 (Q9) carbon atoms (Figure 29.10). The predominant ubiquinone in humans is Qio while in rodents it is Q9. Ubiquinones are especially abundant in the mitochondrial respiratory chain where their concentration is about 100 times higher than that of other electron carriers. Ubihydroquinone Q10 is also found in LDL where it supposedly exhibits the antioxidant activity (see Chapter 23). 

Abnormalities of the respiratoiy chain. These are increasingly identified as the hallmark of mitochondrial diseases or mitochondrial encephalomyopathies [13]. They can be identified on the basis of polarographic studies showing differential impairment in the ability of isolated intact mitochondria to use different substrates. For example, defective respiration with NAD-dependent substrates, such as pyruvate and malate, but normal respiration with FAD-dependent substrates, such as succinate, suggests an isolated defect of complex I (Fig. 42-3). However, defective respiration with both types of substrates in the presence of normal cytochrome c oxidase activity, also termed complex IV, localizes the lesions to complex III (Fig. 42-3). Because frozen muscle is much more commonly available than fresh tissue, electron transport is usually measured through discrete portions of the respiratory chain. Thus, isolated defects of NADH-cytochrome c reductase, or NADH-coenzyme Q (CoQ) reductase suggest a problem within complex I, while a simultaneous defect of NADH and succinate-cytochrome c reductase activities points to a biochemical error in complex III (Fig. 42-3). Isolated defects of complex III can be confirmed by measuring reduced CoQ-cytochrome c reductase activity. 

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