Erythrocytes from individuals with pyruvate kinase (PK) deficiency have approximately half the total (oxidized and reduced) nicotinamide adenine dinucleotide (NAD) of normal erythrocytes. In order to elucidate the mechanism(s) for the decrease in total NAD, we examined NAD synthesis in intact erythrocytes. It is demonstrated that NAD synthesis is impaired in PK-deficient erythrocytes to a degre...

Tumor nicotinamide adenine dinudeotide (NAD) glycohydrolase (EC.3.2.2.5) was purified 500-fold, and this preparation was used to study the influence of NAD glycohydrolase on the NAD + content and activity of crystalline glyceraldehyde-3phosphate dehydrogenase. The pseudomonomolecular velocity constant, k, for the hydrolysis of free NAD + was 5-fold that of the constant for the enzyme-bound NAD ...

The Sir2 protein is an NAD(+)-dependent protein deacetylase that is required for silencing at the silent mating-type loci, telomeres, and the ribosomal DNA (rDNA). Mutations in the NAD(+) salvage gene NPT1 weaken all three forms of silencing and also cause a reduction in the intracellular NAD(+) level. We now show that mutation of a highly conserved histidine residue in Npt1p results in a silen...

Tumor nicotinamide adenine dinudeotide (NAD) glycohydrolase (EC.3.2.2.5) was purified 500-fold, and this preparation was used to study the influence of NAD glycohydrolase on the NAD + content and activity of crystalline glyceraldehyde-3phosphate dehydrogenase. The pseudomonomolecular velocity constant, k, for the hydrolysis of free NAD + was 5-fold that of the constant for the enzyme-bound NAD ...

The increase in the frequency of drug resistance bacterial infections has led to research new antibacterial agents targeting mechanisms. Many functions NAD+-dependent DNA ligase have made it a remarkable target for discovery. Escherichia coli (E. coli) is presented as potential due its unique substrate specificity compared ATP-dependent human ligase. In this study, was aimed produce and purify ...

NQO1 is a FAD containing NAD(P)H-dependent oxidoreductase that catalyzes the reduction of quinones and related substrates. In cells, NQO1 participates in a number of binding interactions with other proteins and mRNA and these interactions may be influenced by the concentrations of reduced pyridine nucleotides. NAD(P)H can protect NQO1 from proteolytic digestion suggesting that binding of reduce...

Hypoacetylated histones are a hallmark of heterochromatin in organisms ranging from yeast to humans. Histone deacetylation is carried out by both NAD(+)-dependent and NAD(+)-independent enzymes. In the budding yeast Saccharomyces cerevisiae, deacetylation of histones in heterochromatic chromosomal domains requires Sir2, a phylogenetically conserved NAD(+)-dependent deacetylase. In the fission y...

The isocitrate dehydrogenase (MfIDH) with unique double coenzyme specificity from Methylobacillus flagellatus was purified and characterized, and its gene was cloned and overexpressed in E. coli as a fused protein. This enzyme is homodimeric,-with a subunit molecular mass of 45 kDa and a specific activity of 182 U mg -1 with NAD+ and 63 U mg -1 with NADP+. The MfIDH activity was dependent on di...

Sirtuin proteins comprise a unique class of NAD+-dependent protein deacetylases. Although several structures of sirtuins have been determined, the mechanism by which NAD+ cleavage occurs has remained unclear. We report the structures of ternary complexes containing NAD+ and acetylated peptide bound to the bacterial sirtuin Sir2Tm and to a catalytic mutant (Sir2Tm(H116Y)). NAD+ in these structur...

To characterize novel signaling pathways that underlie NAD(P)H oxidase-mediated signaling in atherosclerosis, we first examined differences in thrombin-induced gene expression between wild-type and p47phox(-/-) (NAD[P]H oxidase-deficient) VSMC. Of the 9000 genes analyzed by cDNA microarray method at the G1/S transition point, 76 genes were similarly and significantly modulated in both the cell ...