title: Mass spectrometry-based proteomic analysis of human liver cytochrome(s) P450.
abstract: The major objective of personalized medicine is to select optimized drug therapies and to a large degree such mission is determined by the expression profiles of cytochrome(s) P450 (CYP). Accordingly, a proteomic case study in personalized medicine is provided by the superfamily of cytochromes P450. Our knowledge about CYP isozyme expression on a protein level is very limited and based exclusively on DNA/mRNA derived data. Such information is not sufficient because transcription and translation events do not lead to correlated levels of expressed proteins. Here we report expression profiles of CYPs in human liver obtained by mass spectrometry (MS)-based proteomic approach. We analyzed 32 samples of human liver microsomes (HLM) of different sexes, ages and ethnicity along with samples of recombinant human CYPs. We have experimentally confirmed that each CYP isozyme can be effectively differentiated by their unique isozyme-specific tryptic peptide(s). Trypsin digestion patterns for almost 30 human CYP isozymes were established. Those findings should assist in selecting tryptic peptides suitable for MS-based quantitation. The data obtained demonstrate remarkable differences in CYP expression profiles. CYP2E1, CYP2C8 and CYP4A11 were the only isozymes found in all HLM samples. Female and pediatric HLM samples revealed much more diverse spectrum of expressed CYPs isozymes compared to male HLM. We have confirmed expression of a number of "rare" CYP (CYP2J2, CYP4B1, CYP4V2, CYP4F3, CYP4F11, CYP8B1, CYP19A1, CYP24A1 and CYP27A1) and obtained first direct experimental data showing expression of such CYPs as CYP2F1, CYP2S1, CYP2W1, CYP4A22, CYP4X1, and CYP26A1 on a protein level.
authors.affiliation: Tumor Vaccines and Biotechnology Branch, Office of Cellular, Tissue and Gene Therapies, Center for Biologics Evaluation and Research, FDA, Bethesda, MD, USA.
authors.forename: Kamlesh
authors.initials: K
authors.lastname: Shrivas
authors.forename: Samuel T
authors.initials: ST
authors.lastname: Mindaye
authors.forename: Melkamu
authors.initials: M
authors.lastname: Getie-Kebtie
authors.forename: Michail A
authors.initials: MA
authors.lastname: Alterman
mesh_terms: D000595:Amino Acid Sequence; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis* / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods*
chemical_list: D003577:Cytochrome P-450 Enzyme System