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 |
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Authors: | 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 |
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Institution: | Department of Biochemistry, V.P. Chest Institute, University of Delhi, India. |
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Abstract: | 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. |
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Keywords: | Acetyl donor Protein acetylation Matrix-assisted laser desorption/ionization-time of flight Transacetylase |
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