Identification of yacE (coaE) as the structural gene for dephosphocoenzyme A kinase in Escherichia coli K-12.
Dephosphocoenzyme A (dephospho-CoA) kinase catalyzes the final step in coenzyme A biosynthesis, the phosphorylation of the 3'-hydroxy group of the ribose sugar moiety. Wild-type dephospho-CoA kinase from Corynebacterium ammoniagenes was purified to homogeneity and subjected to N-terminal sequence analysis. A BLAST search identified a gene from Escherichia coli previously designated yacE encoding a highly homologous protein. Amplification of the gene and overexpression yielded recombinant dephospho-CoA kinase as a 22.6-kDa monomer. Enzyme assay and nuclear magnetic resonance analyses of the product demonstrated that the recombinant enzyme is indeed dephospho-CoA kinase. The activities with adenosine, AMP, and adenosine phosphosulfate were 4 to 8% of the activity with dephospho-CoA. Homologues of the E. coli dephospho-CoA kinase were identified in a diverse range of organisms.
Mishra
P
P
Department of Chemistry, State University at Stony Brook, New York 11794-3400, USA.
Park
P K
PK
Drueckhammer
D G
DG
D000241:Adenosine / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000250:Adenosine Phosphosulfate / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D016415:Sequence Alignment
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000249:Adenosine Monophosphate; D000250:Adenosine Phosphosulfate; D017853:Phosphotransferases (Alcohol Group Acceptor); C020021:dephospho-CoA kinase; D000241:Adenosine