Pyridine nucleotides oxidation

Walker, G. A. and Kilgour, G. L. 1965. Pyridine nucleotide oxidizing enzymes of Lactobacillus casei. II. Oxidase and peroxidase. Arch. Biochem. Biophys. Ill, 534-539. 

Release of Ca from intact rat liver mitochondria can be induced by oxidation of mitochondrial pyridine nucleotides. This was first shown by the group of Lehninger [8] who oxidized mitochondrial pyridine nucleotides with acetoacetate or oxaloacetate at the level of the citric add cycle. Our group [9 used hydroperoxides, known to be produced by the respiratory chain of mitochondria and to be linked to pyridine nucleotides by glutathione peroxidase and reductase, to induce Ca release and oxidation of pyridine nucleotides. Orrenius and co-workers employed [10] menadione (2-methyl-l, 4-naphthoquinone) to induce pyridine nucleotide oxidation and Ca release. The latter... 

Oxidation alone of pyridine nucleotides is not sufficient to induce Ca release. In the presence of ATP, the hydroperoxide-induced pyridine nucleotide oxidation is even accelerated, yet pyridine nucleotide hydrolysis and Ca release are inhibited [11]. Similar observations were made during the menadione-induced Ca release [10]. When liver mitochondria are treated with N-ethyl maleimide to lower intramitochondrial glutathione, both oxidation of pyridine nucleotides and Ca " release are inhibited (S. Baumhuter, C. Richter, unpubl.). Finally, both pyridine nucleotide hydrolysis and Ca release show the same sigmoidal dependence on the mitochondrial Ca load [15]. Thus, there is clear, albeit circumstantial, evidence that pyridine nucleotide hydrolysis and Ca " release are functionally related. The link between the two processes may be protein ADP-ribosylation. 

Ca release from intact rat liver mitochondria is induced by compoimds that oxidize intramitochondrial pyridine nucleotides. Oxidation alone of pyridine nucleotides is not sufficient to cause Ca release. Release is, however, observed when oxidized pyridine nucleotides are hydrolyzed at the N-glycosidic bond linking ADP-ribose and nicotinamide. Both pyridine nucleotide hydrolysis and Ca release are very similarly dependent on the intramitochondrial Ca load. 

FIGURE 18.10 Hydrogen and electrons released in the course of oxidative catabolism are transferred as hydride ions to the pyridine nucleotide, NAD, to form NADH -t- H in dehydrogenase reactions of the type... 

Nicotinamide is an essential part of two important coenzymes nicotinamide adenine dinucleotide (NAD ) and nicotinamide adenine dinucleotide phosphate (NADP ) (Figure 18.19). The reduced forms of these coenzymes are NADH and NADPH. The nieotinamide eoenzymes (also known as pyridine nucleotides) are electron carriers. They play vital roles in a variety of enzyme-catalyzed oxidation-reduction reactions. (NAD is an electron acceptor in oxidative (catabolic) pathways and NADPH is an electron donor in reductive (biosynthetic) pathways.) These reactions involve direct transfer of hydride anion either to NAD(P) or from NAD(P)H. The enzymes that facilitate such... 

Riboflavin was first isolated from whey in 1879 by Blyth, and the structure was determined by Kuhn and coworkers in 1933. For the structure determination, this group isolated 30 mg of pure riboflavin from the whites of about 10,000 eggs. The discovery of the actions of riboflavin in biological systems arose from the work of Otto Warburg in Germany and Hugo Theorell in Sweden, both of whom identified yellow substances bound to a yeast enzyme involved in the oxidation of pyridine nucleotides. Theorell showed that riboflavin 5 -phosphate was the source of the yellow color in this old yellow enzyme. By 1938, Warburg had identified FAD, the second common form of riboflavin, as the coenzyme in D-amino acid oxidase, another yellow protein. Riboflavin deficiencies are not at all common. Humans require only about 2 mg per day, and the vitamin is prevalent in many foods. This vitamin... 

Xanthobacter sp. strain Py2 may be grown with propene or propene oxide. On the basis of amino acid sequences, the monooxygenase that produces the epoxide was related to those that catalyzes the monooxygenation of benzene and toluene (Zhou et al. 1999). The metabolism of the epoxide is initiated by nucleophilic reaction with coenzyme M followed by dehydrogenation (Eigure 7.13a). There are alternative reactions, both of which are dependent on a pyridine nucleotide-disulfide oxidoreductase (Swaving et al. 1996 Nocek et al. 2002) ... 

Nitrosoarenes are readily formed by the oxidation of primary N-hydroxy arylamines and several mechanisms appear to be involved. These include 1) the metal-catalyzed oxidation/reduction to nitrosoarenes, azoxyarenes and arylamines (144) 2) the 02-dependent, metal-catalyzed oxidation to nitrosoarenes (145) 3) the 02-dependent, hemoglobin-mediated co-oxidation to nitrosoarenes and methe-moglobin (146) and 4) the 0 2-dependent conversion of N-hydroxy arylamines to nitrosoarenes, nitrosophenols and nitroarenes (147,148). Each of these processes can involve intermediate nitroxide radicals, superoxide anion radicals, hydrogen peroxide and hydroxyl radicals, all of which have been observed in model systems (149,151). Although these radicals are electrophilic and have been suggested to result in DNA damage (151,152), a causal relationship has not yet been established. Nitrosoarenes, on the other hand, are readily formed in in vitro metabolic incubations (2,153) and have been shown to react covalently with lipids (154), proteins (28,155) and GSH (17,156-159). Nitrosoarenes are also readily reduced to N-hydroxy arylamines by ascorbic acid (17,160) and by reduced pyridine nucleotides (9,161). 

The most important coenzymes in synthetic organic chemistry [14] and industrially applied biotransformations [15] are the nicotinamide cofactors NAD/ H (3a/8a, Scheme 43.1) and NAD(P)/H (3b/8b, Scheme 43.1). These pyridine nucleotides are essential components of the cell [16]. In all the reactions where they are involved, they serve solely as hydride donors or acceptors. The oxidized and reduced form of the molecules are shown in Scheme 43.1, the redox reaction taking place at the C-4 atom of the nicotinamide moiety. 

Alcohol dehydrogenases catalyze oxidation of alcohols in a reaction dependent on the pyridine nucleotide NAD+ [Eq. (5)]. Since the reaction is reversible, alcohol dehydrogenases also catalyze the reduction of aldehydes by... 

Indicine IV-oxide (169) (Scheme 36) is a clinically important pyrrolizidine alkaloid being used in the treatment of neoplasms. The compound is an attractive drug candidate because it does not have the acute toxicity observed in other pyrrolizidine alkaloids. Indicine IV-oxide apparently demonstrates increased biological activity and toxicity after reduction to the tertiary amine. Duffel and Gillespie (90) demonstrated that horseradish peroxidase catalyzes the reduction of indicine IV-oxide to indicine in an anaerobic reaction requiring a reduced pyridine nucleotide (either NADH or NADPH) and a flavin coenzyme (FMN or FAD). Rat liver microsomes and the 100,000 x g supernatant fraction also catalyze the reduction of the IV-oxide, and cofactor requirements and inhibition characteristics with these enzyme systems are similar to those exhibited by horseradish peroxidase. Sodium azide inhibited the TV-oxide reduction reaction, while aminotriazole did not. With rat liver microsomes, IV-octylamine decreased... 

Isselbacher, K. J., and McCarthy, E. A., The influence of pyridine nucleotides on galactose-1-C14 oxidation to C1402 in vitro. Biochem. Biophys. Research Cotn-muns. 1, 49-53 (1959). 

Figure 4 summarizes the direct oxidative pathway with its relations to (a) the gycolytic route of G-6-P utilization, (b) the reduction of pyridine nucleotides, and (c) the influences on some reactions depending on reduced pyridine nucleotides. 

The reactions in Eqs. (10) and (11) indicate the links with glucose metabolism, the former to the direct oxidative pathway, the latter to the glycolytic route. The essential link in the reduction of MHb is the generation of reduced pyridine nucleotides (B14, R12, S10). The... 

The ALDs are a subset of the superfamily of medium-chain dehydrogenases/reductases (MDR). They are widely distributed, cytosolic, zinc-containing enzymes that utilize the pyridine nucleotide [NAD(P)+] as the catalytic cofactor to reversibly catalyze the oxidation of alcohols to aldehydes in a variety of substrates. Both endobiotic and xenobiotic alcohols can serve as substrates. Examples include (72) ethanol, retinol, other aliphatic alcohols, lipid peroxidation products, and hydroxysteroids (73). 

The aldehyde dehydrogenases are members of a superfamily of pyridine nucleotide [NAD(P)+]-dependant oxidoreductases that catalyze the oxidation of aldehydes to... 

Flavoprotein dehydrogenases usually accept electrons from reduced pyridine nucleotides and donate them to a suitable electron acceptor. The oxidation-reduction midpoint potential of the FAD of the oxidase has been determined by ESR spectroscopy and shown to be -280 mV. The NADP+/ NADPH redox potential is -320 mV and that of the cytochrome b is -245 mV hence, the flavin is thermodynamically capable of accepting electrons from NADPH and transferring them to cytochrome b. As two electrons are transferred from NADPH, although O2 reduction requires only one electron, the scheme of electron transfer shown in Figure 5.8 has been proposed by Cross and Jones (1991). 

Lynen had studied chemistry in Munich under Wieland his skill as a chemist led to the successful synthesis of a number of fatty acyl CoA derivatives which proved to be substrates in the catabolic pathway. Many of these C=0 or C=C compounds had characteristic UV absorption spectra so that enzyme reactions utilizing them could be followed spectrophotometrically. This technique was also used to identify and monitor the flavoprotein and pyridine nucleotide-dependent steps. Independent evidence for the pathway was provided by Barker, Stadtman and their colleagues using Clostridium kluyveri. Once the outline of the degradation had been proposed the individual steps of the reactions were analyzed very rapidly by Lynen, Green, and Ochoa s groups using in the main acetone-dried powders from mitochondria, which, when extracted with dilute salt solutions, contained all the enzymes of the fatty acid oxidation system. 

The liver alcohol dehydrogenase mentioned in the preceding section has the same pro-R stereospecificity for NAD and ethanol as yeast alcohol dehydrogenase. Furthermore, the oxidation of ethanol by a microsomal oxidizing system, or by catalase and H2O2, likewise proceeds with pro-R stereospecificity for the ethanol77>. The catalase-H2C>2 system is so very different, however, from the pyridine nucleotide dehydrogenase, that one wonders whether the similarity in stereospecificity for ethanol is fortuitous. 

It should be noted, in this connection, that there are pyridine nucleotide dehydrogenases which catalyze redox reactions which must occur in two steps. Hydroxymethylglutaryl CoA reductase (discussed on p. 51) is one example. Another is uridine diphosphate-glucose dehydrogenase, which catalyzes the oxidation of the C—6 of the glucose (i.e., a primary alcohol) to a carboxyl group. In both cases, there are two molecules of pyridine nucleotide required, and the overall reactions are essentially irreversible. The former enzyme, with A stereospecificity for the pyridine nucleotide, catalyzes the reduction of an acyl-CoA group... 

The generalization that the same dehydrogenase has the same stereospecificity, no matter what the source of the enzyme, has been tested now particularly well for malic and lactic dehydrogenases. In fact, one can venture a guess, that pyridine nucleotide dehydrogenases which oxidize a-hydroxycarb oxylic acids at the a-position, all have A stereospecificity for the pyridine nucleotide, regardless of their stereo-specificity for the substrate. Biellman and Rosenheimer 88> have assembled the data. One can add liver malic enzyme 90> to their list. 

Intracellular reduced pyridine nucleotides NAD(P)H are the primary suppliers of reducing power to anabolic and catabolic pathways. They can be measured because of their fluorescent properties.<16) The fluorescence is caused by the presence of the reduced forms of the pyridine nucleotides NADH and NADPH (jointly referred to as NAD(P)H). These fluorophores absorb light in a wide band around 340 nm, and reemit, or fluoresce, light in a wide band around 460 nm. The phosphorylated and nonphos-phorylated nucleotides have essentially equivalent fluorescence properties while the oxidized forms of these nucleotides are nonfluorescent. 

It was known that the intracellular concentrations of the reduced and oxidized forms of the pyridine nucleotides vary in different cell types and under different cell culture conditions.(17) Harrison and Chance applied the NAD(P)H fluorescence technique and found that culture fluorescence can be related to the metabolic state of the cells. 18,19) Since then, more than a hundred papers on NAD(P)H fluorometry have been published. However, they are primarily divided into three major categories ... 

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