PMID: 8605288Dec 1, 1995Paper

Purification, mass spectrometric characterization, and covalent modification of murine glutathione S-transferases

Chemical Research in Toxicology
Alyson E MitchellA D Jones

Abstract

Three cytosolic glutathione S-transferase (GST, EC 2.5.1.18) isozymes were purified from livers of male CD-1 mice to homogeneity using affinity chromatography and gradient elution. Isozyme molecular masses and purities were determined using electrospray ionization mass spectrometry (ESI/MS), HPLC, and electrophoretic methods. Isozymes were assigned to alpha, pi, and mu classes based on kinetic and electrophoretic properties. Molecular masses determined by mass spectrometry were 25 271.4 +/- 2, 23 478.8 +/- 2, and 25 838.4 +/- 2 Da for alpha, pi, and mu isozymes, respectively. Molecular masses for pi and mu class isozymes agreed with molecular masses derived from nucleic acid sequences, but the molecular mass of the alpha class isozyme was 42 Da greater than calculated for a reported sequence. Analysis of tryptic digests of GST-alpha using tandem mass spectrometry determined that the molecular weight discrepancy could be attributed to N-terminal acetylation. Effects of covalent attachment of 4-hydroxy-2-nonenal (HNE) on catalytic activities of each GST isozyme were investigated by incubating GSTs with HNE. The extent of HNE-protein adduct formation was determined using ESI/MS. Catalytic activity of HNE-modified GSTs toward CDNB ...Continue Reading

Citations

Oct 12, 2001·Archives of Biochemistry and Biophysics·P S WongA van der Vliet
May 20, 2008·Archives of Toxicology·Jaime L RudolfAlyson E Mitchell
May 12, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Munetaka OzekiShinya Toyokuni
Apr 12, 2003·Progress in Lipid Research·Koji Uchida
Mar 23, 2002·Protein Science : a Publication of the Protein Society·John W CrabbHenry F Hoff
Mar 16, 2011·Drug Metabolism Reviews·Larissa M Balogh, William M Atkins
Nov 27, 2007·Free Radical Biology & Medicine·Yukihiro TsuchiyaHiroshi Hojo
Jan 11, 2011·Chemico-biological Interactions·Jean-François LesgardsPierre Stocker
May 11, 2004·Mass Spectrometry Reviews·Marina CariniRoberto Maffei Facino
Oct 18, 2006·Medicinal Research Reviews·Giancarlo AldiniMarina Carini
Nov 30, 2019·Oxidative Medicine and Cellular Longevity·Himangshu Sonowal, Kota V Ramana
Sep 26, 2007·Chemical Research in Toxicology·Alexandra DruckovaLawrence J Marnett

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