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题名: SDS-PAGE-free protocol for comprehensive identification of cytochrome P450 enzymes and uridine diphosphoglucuronosyl transferases in human liver microsomes
作者: Sun, Liangliang1;  Zhang, Yanyan2;  Tao, Dingyin1;  Zhu, Guijie1;  Zhao, Qun1, 3;  Wu, Qi1, 3;  Liang, Zhen1;  Yang, Ling2;  Zhang, Lihua1;  Zhang, Yukui1
关键词: 1-Butyl-3-methyl imidazolium tetrafluoroborate ;  2D-LC-ESI-MS/MS ;  Cytochrome P450 enzyme ;  Protein spectral count ;  Uridine diphosphoglucuronosyl transferase
刊名: PROTEOMICS
发表日期: 2012-12-01
DOI: 10.1002/pmic.201200260
卷: 12, 期:23-24, 页:3464-3469
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ;  Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: Comprehensive identification of cytochrome P450 enzymes (CYPs) and uridine diphosphoglucuronosyl transferases (UGTs) in human liver microsomes (HLMs) was performed with an SDS-PAGE-free protocol. HLMs were solubilized with 5% v/v ionic liquid, 1-butyl-3-methyl imidazolium tetrafluoroborate, followed by tryptic digestion, and 2D-SCX-RPLC-ESI-MS/MS (LTQ XL) analysis in triplicate. In total, 27 CYPs and 12 UGTs were confidently identified with average sequence coverage as 30.99 and 25.07%, average peptide number as 14 and 13, and average unique peptide number as 7 and 4, respectively. The highly similar isoforms of CYP3A, CYP2C, and CYP4F subfamilies could be unambiguously differentiated from each other, despite the fact that the sequence similarity of CYP2C9 and CYP2C19 is 91%. In addition, protein spectral count was used to approximately evaluate the relative abundance of identified CYPs and UGTs, and the results agreed with previous immunochemistry reports.
关键词[WOS]: TANDEM MASS-SPECTROMETRY ;  PROTEOMIC APPROACH ;  ISOFORMS ;  P450 ;  QUANTIFICATION ;  THERAPEUTICS ;  EXPRESSION ;  TISSUE
语种: 英语
WOS记录号: WOS:000312650100003
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/143063
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Lab High Efficient Separat & High Sensit Characte, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Lab Pharmaceut Resource Discovery, Dalian 116023, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China

Recommended Citation:
Sun, Liangliang,Zhang, Yanyan,Tao, Dingyin,et al. SDS-PAGE-free protocol for comprehensive identification of cytochrome P450 enzymes and uridine diphosphoglucuronosyl transferases in human liver microsomes[J]. PROTEOMICS,2012,12(23-24):3464-3469.
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