The Pur10 protein encoded in the gene cluster for puromycin biosynthesis of Streptomyces alboniger is an NAD-dependent ATP dehydrogenase.
The pur10 gene of the puromycin (pur) cluster of Streptomyces alboniger is essential for the biosynthesis of this antibiotic. Highly purified Pur10 protein, obtained in Escherichia coli as a recombinant protein fused to a histidine tail, had an NAD-dependent ATP dehydrogenase activity. The Km and Vmax values for ATP were 0.49 mM and 14.5 nmol/min and for NAD 0.53 mM and 15.2 nmol/min, respectively. The ATP-derived product of the reaction apparently decomposed producing a triphosphorylated compound plus an adenine derivative. These and previous results suggested that Pur10 carries out the first step of the puromycin biosynthetic pathway, namely, conversion of ATP into 3'-keto-3'-deoxyATP.
Rubio
M A
MA
Centro de Biologia Molecular Severo Ochoa (CSIC/UAM), Universidad Autónoma, Cantoblanco, Madrid, Spain.
Espinosa
J C
JC
Tercero
J A
JA
Jiménez
A
A
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins; D002384:Catalysis; D004789:Enzyme Activation / Q000235:genetics; D005798:Genes, Bacterial; D020043:Genes, Essential; D007700:Kinetics; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D009243:NAD / Q000378:metabolism*; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011691:Puromycin / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D009243:NAD; D011691:Puromycin; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D000429:Alcohol Oxidoreductases; C115699:Pur10 protein, Streptomyces alboniger