Targeted label-free approach for quantification of epoxide hydrolase and glutathione transferases in microsomes

Analytical Biochemistry
Wei SongZhihong Peng

Abstract

The aim of this study was to investigate the expression and organ distribution of cytochrome P450 (CYP450) enzymes, microsomal epoxide hydrolase (MEH), and microsomal glutathione-S-transferase (MGST 1, 2, 3) in human liver, lung, intestinal, and kidney microsomes by targeted peptide-based quantification using nano liquid chromatography-tandem multiple reaction monitoring (nano LC-MRM). Applying this method, we analyzed 16 human liver microsomes and pooled lung, kidney, and intestine microsomes. Nine of the CYP450s (CYP1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4, 3A5) could be quantified in liver. Except for CYP3A4 and 3A5 existing in intestine, other CYP450s had little content (<0.1 pmol/mg protein) in extrahepatic tissues. MEH and MGSTs could be quantified both in hepatic and in extrahepatic tissues. The highest concentrations of MEH and MGST 1, 2 were found in liver; conversely MGST 3 was abundant in human kidney and intestine compared to liver. The targeted proteomics assay described here can be broadly and efficiently utilized as a tool for investigating the targeted proteins. The method also provides novel CYP450s, MEH, and MGSTs expression data in human hepatic and extrahepatic tissues that will benefit rational approaches to...Continue Reading

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Citations

Jun 15, 2016·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Robert S Foti, Deepak K Dalvie
Jul 12, 2017·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·Guiyuan HeLing Yang
Jun 11, 2017·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Francesca ToselliMartin A Hayes
Mar 9, 2017·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Daniel ScotcherAleksandra Galetin
Dec 20, 2020·European Journal of Drug Metabolism and Pharmacokinetics·Michael J DoerksenAbby C Collier

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