RRC ID 18506
著者 Miura T, Kakehashi H, Shinkai Y, Egara Y, Hirose R, Cho AK, Kumagai Y.
タイトル GSH-mediated S-transarylation of a quinone glyceraldehyde-3-phosphate dehydrogenase conjugate.
ジャーナル Chem Res Toxicol
Abstract Many cellular proteins with reactive thiols form covalent bonds with electrophiles, thereby modifying their structures and activities. Here, we describe the recovery of a glycolytic protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), from such an electrophilic attack by 1,2-napthoquinone (1,2-NQ). GAPDH readily formed a covalent bond with 1,2-NQ through Cys152 at a low concentration (0.2 μM) in a cell-free system, but when human epithelial A549 cells were exposed to this quinone at 20 μM, only minimal binding was observed although extensive binding to numerous other cellular proteins occurred. Depletion of cellular glutathione (GSH) with buthionine sulfoximine (BSO) resulted in some covalent modification of cellular GAPDH by 1,2-NQ and a significant reduction of GAPDH activity in the cells. Incubation of native, but not boiled, human GAPDH that had been modified by 1,2-NQ with GSH resulted in a concentration-dependent removal of 1,2-NQ from the GAPDH conjugate, accompanied by partial recovery of lost catalytic activity and formation of a 1,2-NQ-GSH adduct (1,2-NQ-SG). While GAPDH is recognized as a multifunctional protein, our results show that GAPDH also has a unique ability to recover from electrophilic modification by 1,2-NQ through a GSH-dependent S-transarylation reaction.
巻・号 24(11)
ページ 1836-44
公開日 2011-11-21
DOI 10.1021/tx200025y
PMID 21827172
MeSH Buthionine Sulfoximine / adverse effects Buthionine Sulfoximine / pharmacology Cell Line Cell-Free System Cloning, Molecular Epithelial Cells / drug effects Epithelial Cells / enzymology* Escherichia coli Glutathione / metabolism* Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry Glyceraldehyde-3-Phosphate Dehydrogenases / genetics Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism* Glycolysis / drug effects Glycolysis / genetics Humans Mutation Naphthoquinones / chemistry Naphthoquinones / metabolism* Oxidation-Reduction / drug effects Plasmids Protein Denaturation Protein Processing, Post-Translational Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism* Sulfhydryl Compounds / chemistry Sulfhydryl Compounds / metabolism* Transformation, Bacterial
IF 3.184
引用数 21
WOS 分野 CHEMISTRY, MEDICINAL TOXICOLOGY CHEMISTRY, MULTIDISCIPLINARY
リソース情報
ヒト・動物細胞 A549(RCB0098)