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• W2011197100 endingPage "6406" @default.
• W2011197100 startingPage "6392" @default.
• W2011197100 abstract "Group VIA phospholipase A2 (iPLA2β) is expressed in phagocytes, vascular cells, pancreatic islet β-cells, neurons, and other cells and plays roles in transcriptional regulation, cell proliferation, apoptosis, secretion, and other events. A bromoenol lactone (BEL) suicide substrate used to study iPLA2β functions inactivates iPLA2β by alkylating Cys thiols. Because thiol redox reactions are important in signaling and some cells that express iPLA2β produce biological oxidants, iPLA2β might be subject to redox regulation. We report that biological concentrations of H2O2, NO, and HOCl inactivate iPLA2β, and this can be partially reversed by dithiothreitol (DTT). Oxidant-treated iPLA2β modifications were studied by LC−MS/MS analyses of tryptic digests and included DTT-reversible events, e.g., formation of disulfide bonds and sulfenic acids, and others not so reversed, e.g., formation of sulfonic acids, Trp oxides, and Met sulfoxides. W460 oxidation could cause irreversible inactivation because it is near the lipase consensus sequence (463GTSTG467), and site-directed mutagenesis of W460 yields active mutant enzymes that exhibit no DTT-irreversible oxidative inactivation. Cys651-sulfenic acid formation could be one DTT-reversible inactivation event because Cys651 modification correlates closely with activity loss and its mutagenesis reduces sensitivity to inhibition. Intermolecular disulfide bond formation might also cause reversible inactivation because oxidant-treated iPLA2β contains DTT-reducible oligomers, and oligomerization occurs with time- and temperature-dependent iPLA2β inactivation that is attenuated by DTT or ATP. Subjecting insulinoma cells to oxidative stress induces iPLA2β oligomerization, loss of activity, and subcellular redistribution and reduces the rate of release of arachidonate from phospholipids. These findings raise the possibility that redox reactions affect iPLA2β functions." @default.
• W2011197100 created "2016-06-24" @default.
• W2011197100 creator A5015602933 @default.
• W2011197100 creator A5016384271 @default.
• W2011197100 creator A5029423133 @default.
• W2011197100 creator A5060184457 @default.
• W2011197100 date "2006-04-28" @default.
• W2011197100 modified "2023-09-27" @default.
• W2011197100 title "Effects of Biological Oxidants on the Catalytic Activity and Structure of Group VIA Phospholipase A₂" @default.
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