Oxidative phosphorylation formation

Bithionol interferes with the neuromuscular physiology of helminths, impairs egg formation, and may cause defects in the protective cuticle covering the worm. At the biochemical level, the oxidative phosphorylation of the worm is inhibited. 

Apart from gastropods, harmful effects of TBT have also been demonstrated in oysters (Environmental Health Criteria 116, Thain and Waldock 1986). Early work established that adult Pacific oysters (Crassostrea gigas) showed shell thickening caused by the development of gel centers when exposed to 0.2 pg/L of TBT fluoride (Alzieu et al. 1982). Subsequent work established the no observable effect level (NOEL) for shell thickening in this, the most sensitive of the tested species, at about 20 ng/L. It has been suggested that shell thickening is a consequence of the effect of TBT on mitochondrial oxidative phosphorylation (Alzieu et al. 1982). Reduced ATP production may retard the function of Ca++ ATPase, which is responsible for the Ca++ transport that leads to CaCOj deposition during the course of shell formation. Abnormal calcification causes distortion of the shell layers. 

Glucose is metabolized to pyruvate by the pathway of glycolysis, which can occur anaerobically (in the absence of oxygen), when the end product is lactate. Aerobic tissues metabolize pyruvate to acetyl-CoA, which can enter the citric acid cycle for complete oxidation to CO2 and HjO, linked to the formation of ATP in the process of oxidative phosphorylation (Figure 16-2). Glucose is the major fuel of most tissues. 

Pathways are compartmentalized within the cell. Glycolysis, glycogenesis, glycogenolysis, the pentose phosphate pathway, and fipogenesis occur in the cytosol. The mitochondrion contains the enzymes of the citric acid cycle, P-oxidation of fatty acids, and of oxidative phosphorylation. The endoplasmic reticulum also contains the enzymes for many other processes, including protein synthesis, glycerofipid formation, and dmg metabolism. 

The reduced coenzymes are oxidized by the respiratory chain linked to formation of ATP. Thus, the cycle is the major route for the generation of ATP and is located in the matrix of mitochondria adjacent to the enzymes of the respiratory chain and oxidative phosphorylation. 

In a chemical model for mitrochondrial oxidative phosphorylation/ it has been proposed that the mitochondrial membrane, to which ATP and inorganic phosphate are attached, is held in an extended inactive form (50) by coulombic repulsion of positive charges. On reduction of the membrane by NADH one positive centre is removed, and folding of the membrane can occur with extrusion of water. This creates a non-aqueous environment around the ADP (51) and a metal ion can now catalyse the formation... 

The individual steps of the multistep chemical reduction of COj with the aid of NADPHj require an energy supply. This supply is secured by participation of ATP molecules in these steps. The chloroplasts of plants contain few mitochondria. Hence, the ATP molecules are formed in plants not by oxidative phosphorylation of ADP but by a phosphorylation reaction coupled with the individual steps of the photosynthesis reaction, particularly with the steps in the transition from PSII to PSI. The mechanism of ATP synthesis evidently is similar to the electrochemical mechanism involved in their formation by oxidative phosphorylation owing to concentration gradients of the hydrogen ions between the two sides of internal chloroplast membranes, a certain membrane potential develops on account of which the ATP can be synthesized from ADP. Three molecules of ATP are involved in the reaction per molecule of COj. 

Dinitrophenol is a member of the aromatic family of pesticides, many of which exhibit insecticide and fungicide activity. DNP is considered to be highly toxic to humans, with a lethal oral dose of 14 to 43mg/kg. Environmental exposure to DNP occurs primarily from pesticide runoff to water. DNP is used as a pesticide, wood preservative, and in the manufacture of dyes. DNP is an uncoupler, or has the ability to separate the flow of electrons and the pumping of ions for ATP synthesis. This means that the energy from electron transfer cannot be used for ATP synthesis [75,77]. The mechanism of action of DNP is believed to inhibit the formation of ATP by uncoupling oxidative phosphorylation. 

Recent development of mitochondrial theory of aging is so-called reductive hotspot hypothesis. De Grey [465] proposed that the cells with suppressed oxidative phosphorylation survive by reducing dioxygen at the plasma membrane rather than at the mitochondrial inner membrane. Plasma membrane redox system is apparently an origin of the conversion of superoxide into hydroxyl and peroxyl radicals and LDL oxidation. Morre et al. [466] suggested that plasma membrane oxidoreductase links the accumulation of lesions in mitochondrial DNA to the formation of reactive oxygen species on the cell surface. 

By the mid-1950s, therefore, it had become clear that oxidation in the tricarboxylic acid cycle yielded ATP. The steps had also been identified in the electron transport chain where this apparently took place. Most biochemists expected oxidative phosphorylation would occur analogously to substrate level phosphorylation, a view that was tenaciously and acrimoniously defended. Most hypotheses entailed the formation of some high-energy intermediate X Y which, in the presence of ADP and P( would release X and Y and yield ATP. A formulation of the chemical coupling hypothesis was introduced by Slater in 1953,... 

The spatial separation between the components of the electron transport chain and the site of ATP synthesis was incompatible with simple interpretations of the chemical coupling hypothesis. In 1964, Paul Boyer suggested that conformational changes in components in the electron transport system consequent to electron transfer might be coupled to ATP formation, the conformational coupling hypothesis. No evidence for direct association has been forthcoming but conformational changes in the subunits of the FI particle are now included in the current mechanism for oxidative phosphorylation. 

Figure 22.17 Summary of mechanisms to maintain the ATP/ADP concentration ratio in hypoxic myocardium. A decrease in the ATP/ADP concentration ratio increases the concentrations of AMP and phosphate, which stimulate conversion of glycogen/ glucose to lactic acid and hence ATP generation from glycolysis. The changes also increase the activity of AMP deaminase, which increases the formation and hence the concentration of adenosine. The latter has two major effects, (i) It relaxes smooth muscle in the arterioles, which results in vasodilation that provides more oxygen for aerobic ATP generation (oxidative phosphorylation). (ii) It results in decreased work by the heart (i.e. decrease in contractile activity), (mechanisms given in the text) which decreases ATP utilisation.

In the preceding sections the conversion of purines and purine nucleosides to purine nucleoside monophosphates has been discussed. The monophosphates of adenosine and guanosine must be converted to their di- and triphosphates for polymerization to RNA, for reduction to 2 -deoxyribonucleoside diphosphates, and for the many other reactions in which they take part. Adenosine triphosphate is produced by oxidative phosphorylation and by transfer of phosphate from 1,3-diphosphoglycerate and phosphopyruvate to adenosine diphosphate. A series of transphosphorylations distributes phosphate from adenosine triphosphate to all of the other nucleotides. Two classes of enzymes, termed nucleoside mono-phosphokinases and nucleoside diphosphokinases, catalyse the formation of the nucleoside di- and triphosphates by the transfer of the terminal phosphoryl group from adenosine triphosphate. Muscle adenylate kinase (myokinase)... 

Experiments on the physiological and biochemical mechanisms of action suggest that bromethalin uncouples oxidative phosphorylation in central nervous system mitocondria(19). This could lead to a decreased production of ATP, a diminished activity of Na /K ATPase, and a subsequent fluid build up manifested by fluid-filled vacuoles between the myelin sheaths. This vacuole formation in turn leads to an increased cerebrospinal fluid pressure and increased pressure on the nerve axons, yielding a decrease in nerve impulse, paralysis, and death. 

A measure of oxidative phosphorylation, equal to the ratio of the number phosphate groups esterified (i.e., ATP formation from ADP and phosphate) relative to the atoms of oxygen consumed by the mitochondria. 

Oxidizible substrates from glycolysis, fatty acid or protein catabolism enter the mitochondrion in the form of acetyl-CoA, or as other intermediaries of the Krebs cycle, which resides within the mitochondrial matrix. Reducing equivalents in the form of NADH and FADH pass electrons to complex I (NADH-ubiquinone oxidore-ductase) or complex II (succinate dehydrogenase) of the electron transport chain, respectively. Electrons pass from complex I and II to complex III (ubiquinol-cyto-chrome c oxidoreductase) and then to complex IV (cytochrome c oxidase) which accumulates four electrons and then tetravalently reduces O2 to water. Protons are pumped into the inner membrane space at complexes I, II and IV and then diffuse down their concentration gradient through complex V (FoFi-ATPase), where their potential energy is captured in the form of ATP. In this way, ATP formation is coupled to electron transport and the formation of water, a process termed oxidative phosphorylation (OXPHOS). 

Although the citric acid cycle directly generates only one ATP per turn (in the conversion of succinyl-CoA to succinate), the four oxidation steps in the cycle provide a large flow of electrons into the respiratory chain via NADH and FADH2 and thus lead to formation of a large number of ATP molecules during oxidative phosphorylation. 

We saw in Chapter 14 that the energy yield from the production of two molecules of pyruvate from one molecule of glucose in glycolysis is 2 ATP and 2 NADH. In oxidative phosphorylation (Chapter 19), passage of two electrons from NADH to 02 drives the formation of about 2.5 ATP, and passage of two electrons from FADH2 to 02 yields about 1.5 ATP. This stoichiometry allows us to calculate the overall yield of ATP from the complete... 

TABLE 16-1 Stoichiometry of Coenzyme Reduction and ATP Formation in the Aerobic Oxidation of Glucose via Glycolysis, the Pyruvate Dehydrogenase Complex Reaction, the Citric Acid Cycle, and Oxidative Phosphorylation... 

How is a concentration gradient of protons transformed into ATP We have seen that electron transfer releases, and the proton-motive force conserves, more than enough free energy (about 200 lcJ) per mole of electron pairs to drive the formation of a mole of ATP, which requires about 50 kJ (see Box 13-1). Mitochondrial oxidative phosphorylation therefore poses no thermodynamic problem. But what is the chemical mechanism that couples proton flux with phosphorylation ... 

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