Premium
Establishment of the enzymatic protein acetylation independent of acetyl CoA: recombinant glutathione S ‐transferase 3‐3 is acetylated by a novel membrane‐bound transacetylase using 7,8‐diacetoxy‐4‐methyl coumarin as the acetyl donor
Author(s) -
Kohli Ekta,
Gaspari Marco,
Raj Hanumantharao G,
Parmar Virinder S,
van der Greef Jan,
Gupta Garima,
Kumari Ranju,
Prasad Ashok K,
Goel Sanjay,
Pal Giridhari,
Tyagi Yogesh K,
Jain Subhash C,
Ahmad Nizamuddin,
Watterson Arthur C,
Olsen Carl E
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)03445-2
Subject(s) - acetylation , chemistry , enzyme , biochemistry , acetyl coa , polyacrylamide gel electrophoresis , chromatography , gene
The current knowledge on biological protein acetylation is confined to acetyl CoA‐dependent acetylation of protein catalyzed by specific acetyl transferases and the non‐enzymatic acetylation of protein by acetylated xenobiotics such as aspirin. We have discovered a membrane‐bound enzyme catalyzing the transfer of acetyl groups from the acetyl donor 7,8‐diacetoxy‐4‐methyl coumarin (DAMC) to glutathione S ‐transferase 3‐3 (GST3‐3), termed DAMC:protein transacetylase (TAase). The purified enzyme was incubated with recombinant GST3‐3 subunit and DAMC, the modified protein was isolated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) in gel digested with trypsin and the tryptic digest was analyzed by mass spectrometry. The N‐terminus and six lysines, Lys‐51, ‐82, ‐124, ‐181, ‐191 and ‐210, were found to be acetylated. The acetylation of GST3‐3 described above was not observed in the absence of either DAMC or TAase. These results clearly establish the phenomenon of protein acetylation independent of acetyl CoA catalyzed by a hitherto unknown enzyme (TAase) utilizing a certain xenobiotic acetate (DAMC) as the active acetyl donor.