Structural and functional characterization of human NAD kinase.
NADP is essential for biosynthetic pathways, energy, and signal transduction. Its synthesis is catalyzed by NAD kinase. Very little is known about the structure, function, and regulation of this enzyme from multicellular organisms. We identified a human NAD kinase cDNA and the corresponding gene using available database information. A cDNA was amplified from a human fibroblast cDNA library and functionally overexpressed in Escherichia coli. The obtained cDNA, slightly different from that deposited in the database, encodes a protein of 49 kDa. The gene is expressed in most human tissues, but not in skeletal muscle. Human NAD kinase differs considerably from that of prokaryotes by subunit molecular mass (49 kDa vs 30-35 kDa). The catalytically active homotetramer is highly selective for its substrates, NAD and ATP. It did not phosphorylate the nicotinic acid derivative of NAD (NAAD) suggesting that the potent calcium-mobilizing pyridine nucleotide NAADP is synthesized by an alternative route.
Lerner
F
F
Freie Universität Berlin, Institut für Biochemie, Thielallee 63, 14195 Berlin, Germany.
Niere
M
M
Ludwig
A
A
Ziegler
M
M
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000031:analogs & derivatives* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D014018:Tissue Distribution; D014170:Transformation, Genetic
D012333:RNA, Messenger; D011994:Recombinant Proteins; D009243:NAD; C053851:nicotinamide arabinoside adenine dinucleotide; D017853:Phosphotransferases (Alcohol Group Acceptor); C021844:NAD kinase