Molecular cloning and identification of UTR1 of a yeast Saccharomyces cerevisiae as a gene encoding an NAD kinase.
UTR1 of the yeast Saccharomyces cerevisiae was cloned from the genomic DNA by polymerase chain reaction and expressed in Escherichia coli. Characterization of the purified UTR1p revealed that UTR1p is a NAD kinase consisting of six identical subunits with a molecular mass of 60 kDa. UTR1p specifically phosphorylated NAD in the presence of ATP, dATP, or CTP as phosphoryl donors, and was most active at pH 8.0, 30 degrees C. Km values of UTR1p for NAD and ATP were determined to be 0.50 mM and 0.60 mM, respectively.
Kawai
S
S
Research Institute for Food Science, Kyoto University, Uji, 611-0011, Kyoto, Japan.
Suzuki
S
S
Mori
S
S
Murata
K
K
D003001:Cloning, Molecular; D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
D038181:FMN Reductase; D009247:NADH, NADPH Oxidoreductases; C023903:ferric citrate iron reductase; D017853:Phosphotransferases (Alcohol Group Acceptor); C021844:NAD kinase