Extensive protein carbonylation precedes acrolein-mediated cell death in mouse hepatocytes.
Allyl alcohol hepatotoxicity is mediated by an alcohol dehydrogenase-derived biotranformation product, acrolein. This highly reactive alpha,beta-unsaturated aldehyde readily alkylates model proteins in vitro, forming, among other products, Michael addition adducts that possess a free carbonyl group. Whether such damage accompanies acrolein-mediated toxicity in cells is unknown. In this work we established that allyl alcohol toxicity in mouse hepatocytes involves extensive carbonylation of a wide range of proteins, and that the severity of such damage to a subset of 18-50 kDa proteins closely correlated with the degree of cell death. In addition to abolishing cytotoxicity and glutathione depletion, the alcohol dehydrogenase inhibitor 4-methyl pyrazole strongly attenuated protein carbonylation. Conversely, cyanamide, an aldehyde dehydrogenase inhibitor, enhanced cytotoxicity and protein carbonylation. Since protein carbonylation clearly preceded the loss of membrane integrity, it may be associated with the toxic process leading to cell death.
Burcham
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PC
Molecular Toxicology Research Group, Department of Clinical and Experimental Pharmacology, Adelaide University, Adelaide, South Australia 5005, Australia. philip.burcham@adelaide.edu.au
Fontaine
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D000171:Acrolein / Q000633:toxicity*; D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors; D000818:Animals; D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000077604:Fomepizole; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000494:pharmacology; D051379:Mice; D020005:Propanols / Q000633:toxicity; D011506:Proteins / Q000378:metabolism*; D011720:Pyrazoles / Q000633:toxicity
D004791:Enzyme Inhibitors; D020005:Propanols; D011506:Proteins; D011720:Pyrazoles; C006463:allyl alcohol; D000171:Acrolein; D000077604:Fomepizole; D007770:L-Lactate Dehydrogenase; D000444:Aldehyde Dehydrogenase; D005978:Glutathione