Characterization of human liver cytochromes P450 by combining the biochemical and proteomic approaches

Toxicology in Vitro : an International Journal Published in Association with BIBRA
Natalia A PetushkovaAlexander I Archakov

Abstract

Highly purified human liver microsomes were processed by a combination of the biochemical and proteomic methods. Microsomes were purified from the morphologically normal liver tissue obtained from the resected and discarded masses of surrounding liver upon surgical treatment for hemangioma (control) or hepatic metastases arising from colon cancer (pathology). Proteins of each sample were separated by two-dimensional (2-DE) and one-dimensional electrophoresis (1-DE); selected gel regions were excised, in-gel digested and analyzed by matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry. Analysis of collected fingerprints has revealed a total of 13 microsomal membrane proteins involved in the biotransformation of xenobiotics. These were disulfide isomerase, flavine monooxygenase, NADPH-cytochrome P450 reductase and 10 cytochrome P450 forms, namely: CYPs 1B1, 2A6, 2E1, 2C8, 2C9, 2C10, 2D6, 3A4, 4A11, 4F2. These same samples were characterized by the enzymatic assays using the marker substrates for CYPs 1A, 2B, 3A4, 2C and 2E1. Correlations between mass spectrometric data and enzymatic activities were investigated to demonstrate the manner in which the functional and structural aspects of proteomi...Continue Reading

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Citations

Sep 6, 2007·Drug Metabolism Reviews·Tadeja RezenDamjana Rozman
Jul 6, 2010·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Natalia A PetushkovaAlexander I Archakov
Jun 5, 2007·Clinics in Liver Disease·Ancha BaranovaZobair M Younossi
Dec 22, 2012·Journal of Proteome Research·Victor G ZgodaAlexander I Archakov
Mar 10, 2009·Biochemistry. Biokhimii︠a︡·A V LisitsaA I Archakov
Sep 1, 2007·Proteomics. Clinical Applications·Fernando VivancoMaria G Barderas
Jan 29, 2013·Biomedit︠s︡inskai︠a︡ khimii︠a︡·N E Moskaleva, V G Zgoda
Jan 16, 2010·Journal of Proteome Research·Yanping SongFuchu He

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