title: Altered protein S-glutathionylation identifies a potential mechanism of resistance to acetaminophen-induced hepatotoxicity.
abstract: Acetaminophen (APAP) is the most commonly used over-the-counter analgesic. However, hepatotoxicity induced by APAP is a major clinical issue, and the factors that define sensitivity to APAP remain unclear. We have previously demonstrated that mice nulled for glutathione S-transferase Pi (GSTP) are resistant to APAP-induced hepatotoxicity. This study aims to exploit this difference to delineate pathways of importance in APAP toxicity. We used mice nulled for GSTP and heme oxygenase-1 oxidative stress reporter mice, together with a novel nanoflow liquid chromatography-tandem mass spectrometry methodology to investigate the role of oxidative stress, cell signaling, and protein S-glutathionylation in APAP hepatotoxicity. We provide evidence that the sensitivity difference between wild-type and Gstp1/2(-/-) mice is unrelated to the ability of APAP to induce oxidative stress, despite observing significant increases in c-Jun N-terminal kinase and extracellular signal-regulated kinase phosphorylation in wild-type mice. The major difference in response to APAP was in the levels of protein S-glutathionylation: Gstp1/2(-/-) mice exhibited a significant increase in the number of S-glutathionylated proteins compared with wild-type animals. Remarkably, these S-glutathionylated proteins are involved in oxidative phosphorylation, respiratory complexes, drug metabolism, and mitochondrial apoptosis. Furthermore, we found that S-glutathionylation of the rate-limiting glutathione-synthesizing enzyme, glutamate cysteine ligase, was markedly increased in Gstp1/2(-/-) mice in response to APAP. The data demonstrate that S-glutathionylation provides an adaptive response to APAP and, as a consequence, suggest that this is an important determinant in APAP hepatotoxicity. This work identifies potential novel avenues associated with cell survival for the treatment of chemical-induced hepatotoxicity.
authors: Molecular Pharmacology Group, School of Medicine, Jacqui Wood Cancer Centre, University of Dundee, Dundee, United Kingdom (D.J.M., C.R.W., C.J.H.); and Bioinformatics and Biostatistics Group, Cancer Research UK London Research Institute, London, United Kingdom (P.C.).
authors: David J
authors: DJ
authors: McGarry
authors: Molecular Pharmacology Group, School of Medicine, Jacqui Wood Cancer Centre, University of Dundee, Dundee, United Kingdom (D.J.M., C.R.W., C.J.H.); and Bioinformatics and Biostatistics Group, Cancer Research UK London Research Institute, London, United Kingdom (P.C.).
authors: Probir
authors: P
authors: Chakravarty
authors: Molecular Pharmacology Group, School of Medicine, Jacqui Wood Cancer Centre, University of Dundee, Dundee, United Kingdom (D.J.M., C.R.W., C.J.H.); and Bioinformatics and Biostatistics Group, Cancer Research UK London Research Institute, London, United Kingdom (P.C.).
authors: C Roland
authors: CR
authors: Wolf
authors: Molecular Pharmacology Group, School of Medicine, Jacqui Wood Cancer Centre, University of Dundee, Dundee, United Kingdom (D.J.M., C.R.W., C.J.H.); and Bioinformatics and Biostatistics Group, Cancer Research UK London Research Institute, London, United Kingdom (P.C.) c.j.henderson@dundee.ac.uk.
authors: Colin J
authors: CJ
authors: Henderson
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D000700:Analgesics / Q000633:toxicity*; D000818:Animals; D058633:Antipyretics / Q000633:toxicity*; D017209:Apoptosis; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008928:Mitochondria / Q000378:metabolism; D010085:Oxidative Phosphorylation; D018384:Oxidative Stress; D011499:Protein Processing, Post-Translational; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000700:Analgesics; D058633:Antipyretics; D000082:Acetaminophen; D005982:Glutathione Transferase; D048049:Extracellular Signal-Regulated MAP Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D005978:Glutathione