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Establishment of the enzymatic protein acetylation independent of acetyl CoA : recombinant glutathione S-transferase 3-3 is acetylated by a novel membrane-bound transacetylane using 7,8-diacetoxy-4-methyl coumarin as the acetyl donor

Author: Kohli, E. · Gaspari, M. · Raj, H.G. · Parmar, V.S. · Greef, J. van der · Gupta, G. · Kumari, R. · Prasad, A.K. · Goel, S. · Pal, G. · Tyagi, K. · Jain, S.C. · Ahmad, N. · Watterson, A.C. · Olsen, C.E.
Type:article
Date:2002
Institution: Centraal Instituut voor Voedingsonderzoek TNO TNO Voeding
Source:FEBS Letters, 1-3, 530, 139-142
Identifier: 87760
doi: doi:10.1016/S0014-5793(02)03445-2
Keywords: Acetyl Coenzyme A · Acetylation · Acetylesterase · Coumarins · Glutathione Transferase · Recombinant Proteins · Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

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. © 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. Chemicals/CAS: 7,8-diacetoxy-4-methylcoumarin; Acetyl Coenzyme A, 72-89-9; Acetylesterase, EC 3.1.1.6; Coumarins; Glutathione Transferase, EC 2.5.1.18; Recombinant Proteins