Discovery of amide (peptide) bond synthetic activity in Acyl-CoA synthetase.
Acyl-CoA synthetase, which is one of the acid-thiol ligases (EC 6.2.1), plays key roles in metabolic and regulatory processes. This enzyme forms a carbon-sulfur bond in the presence of ATP and Mg(2+), yielding acyl-CoA thioesters from the corresponding free acids and CoA. This enzyme belongs to the superfamily of adenylate-forming enzymes, whose three-dimensional structures are analogous to one another. We here discovered a new reaction while studying the short-chain acyl-CoA synthetase that we recently reported (Hashimoto, Y., Hosaka, H., Oinuma, K., Goda, M., Higashibata, H., and Kobayashi, M. (2005) J. Biol. Chem. 280, 8660-8667). When l-cysteine was used as a substrate instead of CoA, N-acyl-l-cysteine was surprisingly detected as a reaction product. This finding demonstrated that the enzyme formed a carbon-nitrogen bond (EC 6.3.1 acid-ammonia (or amide) ligase (amide synthase); EC 6.3.2 acid-amino acid ligase (peptide synthase)) comprising the amino group of the cysteine and the carboxyl group of the acid. N-Acyl-d-cysteine, N-acyl-dl-homocysteine, and N-acyl-l-cysteine methyl ester were also synthesized from the corresponding cysteine analog substrates by the enzyme. Furthermore, this unexpected enzyme activity was also observed for acetyl-CoA synthetase and firefly luciferase, indicating the generality of the new reaction in the superfamily of adenylate-forming enzymes.
Abe
Tomoko
T
Institute of Applied Biochemistry, and Graduate School of Life and Environmental Sciences, The University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8572, Japan.
Hashimoto
Yoshiteru
Y
Hosaka
Hideaki
H
Tomita-Yokotani
Kaori
K
Kobayashi
Michihiko
M
D000106:Acetate-CoA Ligase / Q000378:metabolism / Q000502:physiology*; D000577:Amides / Q000737:chemistry*; D001671:Biochemistry / Q000379:methods; D002087:Butyrates / Q000737:chemistry; D002244:Carbon / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D008156:Luciferases / Q000378:metabolism; D008274:Magnesium / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D009584:Nitrogen / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D011549:Pseudomonas / Q000201:enzymology; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013455:Sulfur / Q000737:chemistry
D000577:Amides; D002087:Butyrates; D010455:Peptides; D013455:Sulfur; D002244:Carbon; D008156:Luciferases; D000106:Acetate-CoA Ligase; D008274:Magnesium; D003545:Cysteine; D009584:Nitrogen