Identification, molecular cloning and functional characterization of a novel NADH kinase from Arabidopsis thaliana (thale cress).

ABSTRACT: NADH kinase (NADHK; ATP:NADH 2'-phosphotransferase; EC 2.7.1.86), an enzyme that preferentially utilizes NADH as the diphosphonicotinamide nucleotide donor, has been identified for the first time in plants. Low activity (0.4 nmol of NADPH produced/min per mg of protein) was observed in clarified protein extracts from Arabidopsis thaliana (thale cress) cell suspension cultures. However, unlike an NADHK from yeast (Saccharomyces cerevisiae) (POS5), the enzyme from Arabidopsis did not associate with the mitochondria. NADHK was cloned (gi:30699338) from Arabidopsis and studied as a recombinant protein following affinity purification from Escherichia coli. The enzyme had a pH optimum for activity of 7.9 and a subunit molecular mass of 35 kDa. Analytical gel filtration demonstrated that the reco

SUBMITTER: Turner WL 

PROVIDER: S-EPMC1134690 | biostudies-literature | 2005 Jan

REPOSITORIES: biostudies-literature