Nicotinamide adenine dinucleotide substrate oxidation

Since NADPH absorbs light at 340 nm and NADP+ does not (Figure 8-3), this reaction can be followed by measurement of the change in absorbance at this wavelength with time. A similar assay (with lactate and NAD+) can be used for serum lactate dehydrogenase. The difference in absorbance is measured so readily that a number of assays make use of it directly or indirectly. In the latter case, the reaction to be measured is linked by one or more steps to a reaction in which a nicotinamide adenine dinucleotide is oxidized or reduced. Thus, in the measurement of aminotransferases (Chapter 17), one of the products of the primary reaction serves as a substrate for a secondary reaction in which NADH is required. For example, in the assay of aspartate aminotransferase,... 

Glucose [50-99-7] urea [57-13-6] (qv), and cholesterol [57-88-5] (see Steroids) are the substrates most frequentiy measured, although there are many more substrates or metaboUtes that are determined in clinical laboratories using enzymes. Co-enzymes such as adenosine triphosphate [56-65-5] (ATP) and nicotinamide adenine dinucleotide [53-84-9] in its oxidized (NAD" ) or reduced (NADH) [58-68-4] form can be considered substrates. Enzymatic analysis is covered in detail elsewhere (9). 

Reduced nicotinamide-adenine dinucleotide (NADH) plays a vital role in the reduction of oxygen in the respiratory chain [139]. The biological activity of NADH and oxidized nicotinamideadenine dinucleotide (NAD ) is based on the ability of the nicotinamide group to undergo reversible oxidation-reduction reactions, where a hydride equivalent transfers between a pyridine nucleus in the coenzymes and a substrate (Scheme 29a). The prototype of the reaction is formulated by a simple process where a hydride equivalent transfers from an allylic position to an unsaturated bond (Scheme 29b). No bonds form between the n bonds where electrons delocalize or where the frontier orbitals localize. The simplified formula can be compared with the ene reaction of propene (Scheme 29c), where a bond forms between the n bonds. 

The sirtuins (silent information regulator 2-related proteins class III HDACs) form a specific class of histone deacetylases. First, they do not share any sequence or structural homology with the other HDACs. Second, they do not require zinc for activity, but rather use the oxidized form of nicotinamide adenine dinucleotide (NAD ) as cofactor. The reaction catalyzed by these enzymes is the conversion of histones acetylated at specific lysine residues into deacetylated histones, the other products of the reaction being nicotinamide and the metabolite 2 -0-acetyl-adenosine diphosphate ribose (OAADPR) [51, 52]. As HATs and other HDACs, sirtuins not only use acetylated histones as substrates but can also deacetylate other proteins. Intriguingly, some sirtuins do not display any deacetylase activity but act as ADP-ribosyl transferases. 

It is converted to coenzymes, nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP). These coenzymes are bound to hydrogenases, function as oxidants by accepting hydrogen and electrons from substrates and become reduced. 

Functioning of the enzyme requires the presence of a coenzyme, nicotinamide adenine dinucleotide which exists in its oxidized (NAD+) or reduced (NADH) forms. The structure of NADH is shown in (177). Reduction or oxidation occurs by transfer of the pro-R C-4 hydrogen atom of the nicotinamide stereospecifically to or from the substrate. The reaction is therefore a ternary one, with the substrate and coenzyme necessarily within the active site for the reaction to occur.l46Sa... 

Horse liver alcohol dehydrogenase is a well-documented enzyme capable of operating with high stereoselectivity on a broad structural range of alcohol and carbonyl substrates. The present reaction proceeds via the pathway shown below, where NAD and NADH represent the oxidized and reduced forms, respectively, of the nicotinamide adenine dinucleotide coenzyme. 

Methylene-Tetrahydrofolate Reductase The reduction of methylene-tetrahydrofolate to methyl-tetrahydrofolate, shown in Figure 10.7, is catalyzed hy methylene-tetrahydrofolate reductase, a flavin adenine dinucleotide-dependent enzyme during the reaction, the pteridine ring of the substrate is oxidized to dihydrofolate, then reduced to tetrahydrofolate by the flavin, which is reduced by nicotinamide adenine dinucleotide phosphate (NADPH Matthews and Daubner, 1982). The reaction is irreversible under physiological conditions, and methyl-tetrahydrofolate - which is the main form of folate taken up into tissues (Section 10.2.2) - can only be utilized after demethylation catalyzed by methionine synthetase (Section 10.3.4). 

The nature of oxidative processes requires the removal of dectrons from the substrate and many enzymes of the redox class contain transition metals which act as an electron sink." Those enzymes which do not satisfy this requirement need organic cofactors such as nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide 2 -phosphate to act as dectron accqitors, although simple qui-nones have been shown to suffice. ... 

The 3-carbamidopyridinium ring is the chemically active portion of the enzymatic cofactors, NAD and NADP (nicotinamide adenine dinucleotide and its phosphate). A typical reaction involving NAD is the stereospecific (with respect to both cofactor and substrate) oxidation of ethanol to acetaldehyde catalyzed by the enzyme, alcohol dehydrogenase (Eq. 33). 

Enzyme Cofactors. In many enzymatic reactions, and in particular biological reactions, a second substrate (i.e., species) must be introduced to activate the enzyme. This substrate, which is referred to as a cofactor or coenzyme even though it is not an enzyme as such, attaches to the enzyme and is most (often either reduced or oxidized during the course of the reaction. The enzyme-cofac-tor complex is referred to as a holoenzyme. The inactive form of the enzyime-cofactor complex for a specific reaction and reaction direction is called an apoenzyme. An example of the type of system in which a cofactor is used is the formation of ethanol from acetaldehyde in the presence of the enzyme alcohol dehydrogenase (ADH) and the cofactor nicotinamide adenine dinucleotide (NAD) ... 

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