title: Characterization of NADH kinase from Saccharomyces cerevisiae.
abstract: At least two enzymes that phosphorylate diphosphopyridine nucleotides were detected in Saccharomyces cerevisiae: NADH-specific kinase was localized exclusively in the mitochondria, and NAD+-specific kinase was distributed in the microsomal and cytosol fractions but not in the mitochondria. The identity of NAD+ kinase detected in the two fractions remains equivocal. NADH kinase was highly purified 1,041-fold from the mitochondrial fraction. The Km values for NADH and ATP were 105 microM and 2.1 mM, respectively. The relative molecular mass was estimated to be 160,000 by means of molecular sieve chromatography. From inactivation studies with SH inhibitors and protection by NADH, it was demonstrated that a cysteine residue is involved in the binding site of NADH.
authors.affiliation: Department of Agricultural Chemistry, Faculty of Agriculture, Hokkaido University.
authors.forename: Y
authors.initials: Y
authors.lastname: Iwahashi
authors.forename: A
authors.initials: A
authors.lastname: Hitoshio
authors.forename: N
authors.initials: N
authors.lastname: Tajima
authors.forename: T
authors.initials: T
authors.lastname: Nakamura
mesh_terms: D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006882:Hydroxyapatites; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D024101:Mitochondrial Proteins; D008970:Molecular Weight; D010770:Phosphotransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor); D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013104:Spheroplasts / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D006882:Hydroxyapatites; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C021844:NAD kinase; C060670:POS5 protein, S cerevisiae