DICP OpenIR
Subject Area物理化学
Quantitative analysis of cytochrome P450 isoforms in human liver microsomes by the combination of proteomics and chemical probe-based assay
Liu, Xidong1,2; Hu, Lianghai1,2; Ge, Guangbo3; Yang, Bo1,2; Ning, Jing3; Sun, Shixin4; Yang, Ling3; Pors, Klaus5; Gu, Jingkai1,2; XidongLiu; Ge GB(葛广波); JingkaiGu
KeywordBiomedicine Cytochrome P450 Mrm Protein Quantification Stable Isotope Labeling
Source PublicationPROTEOMICS
2014-08-01
DOI10.1002/pmic.201400025
Volume14Issue:16Pages:1943-1951
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordDRUG-DRUG INTERACTIONS ; GENETIC POLYMORPHISMS ; MASS-SPECTROMETRY ; HEPATIC MICROSOMES ; RT-PCR ; CYP3A4 ; METABOLISM ; EXPRESSION ; RAT ; PHARMACOKINETICS
AbstractCytochrome P450 (CYP) is one of the most important drug-metabolizing enzyme families, which participates in the biotransformation of many endogenous and exogenous compounds. Quantitative analysis of CYP expression levels is important when studying the efficacy of new drug molecules and assessing drug-drug interactions in drug development. At present, chemical probe-based assay is the most widely used approach for the evaluation of CYP activity although there are cross-reactions between the isoforms with high sequence homologies. Therefore, quantification of each isozyme is highly desired in regard to meeting the ever-increasing requirements for carrying out pharmacokinetics and personalized medicine in the academic, pharmaceutical, and clinical setting. Herein, an absolute quantification method was employed for the analysis of the seven isoforms CYP1A2, 2B6, 3A4, 3A5, 2C9, 2C19, and 2E1 using a proteome-derived approach in combination with stable isotope dilution assay. The average absolute amount measured from twelve human liver microsomes samples were 39.3, 4.3, 54.0, 4.6, 10.3, 3.0, and 9.3 (pmol/mg protein) for 1A2, 2B6, 3A4, 3A5, 2C9, 2C19, and 2E1, respectively. Importantly, the expression level of CYP3A4 showed high correlation (r = 0.943, p < 0.0001) with the functional activity, which was measured using bufalin-a highly selective chemical probe we have developed. The combination of MRM identification and analysis of the functional activity, as in the case of CYP3A4, provides a protocol which can be extended to other functional enzyme studies with wide application in pharmaceutical research.
Language英语
WOS IDWOS:000340422000010
Citation statistics
Cited Times:23[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/144430
Collection中国科学院大连化学物理研究所
Corresponding AuthorXidongLiu; Ge GB(葛广波); JingkaiGu
Affiliation1.Jilin Univ, Key Lab Mol Enzymol & Engn, Minist Educ, Changchun 130012, Peoples R China
2.Jilin Univ, Res Ctr Drug Metab, Sch Life Sci, Changchun 130012, Peoples R China
3.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian, Peoples R China
4.Appl Biosyst Inc, Asia Pacific Applicat Support Ctr, Shanghai, Peoples R China
5.Univ Bradford, Sch Life Sci, Inst Canc Therapeut, Bradford BD7 1DP, W Yorkshire, England
Recommended Citation
GB/T 7714
Liu, Xidong,Hu, Lianghai,Ge, Guangbo,et al. Quantitative analysis of cytochrome P450 isoforms in human liver microsomes by the combination of proteomics and chemical probe-based assay[J]. PROTEOMICS,2014,14(16):1943-1951.
APA Liu, Xidong.,Hu, Lianghai.,Ge, Guangbo.,Yang, Bo.,Ning, Jing.,...&JingkaiGu.(2014).Quantitative analysis of cytochrome P450 isoforms in human liver microsomes by the combination of proteomics and chemical probe-based assay.PROTEOMICS,14(16),1943-1951.
MLA Liu, Xidong,et al."Quantitative analysis of cytochrome P450 isoforms in human liver microsomes by the combination of proteomics and chemical probe-based assay".PROTEOMICS 14.16(2014):1943-1951.
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