Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach

	Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach
	Liu Xi-Dong 1; Zhu Jun 2; Cong Yu-Ting 1; Hu Liang-Hai 1; Ye Ming-Liang 2; Gu Jing-Kai 1; Zou Han-Fa 2
刊名	chinese journal of analytical chemistry
	2014
卷号	42 期号:1 页码:10-15
关键词	Uridinediphosphate glucuronosyl transferase Multiple reaction monitoring Liver microsome Cytochrome P450 Liquid chromatography-tandem mass spectrometry
英文摘要	strategy for absolute quantification of metabolic enzymes in rat liver microsomes was developed by using shotgun-based proteomic approach. rat liver microsome was digested by trypsin and the specific peptides of drug metabolic enzymes were determined by multiple reaction monitoring (mrm)-based lc-ms/ms. firstly, standard curves were employed for the quantification of p450 and uridinediphosphate glucuronosyl transferase (ugt), which showed good linearity with the r values were all larger than 0.995 and the low limit of quantification was <= 10 nmol/l. secondly, the synthetic isotope labeled specific peptide was served as internal standard for the quantification of ugt1a1. the stable isotope labeled specific peptide showed the same behavior with unlabeled peptide by lc-ms/ms, and the labeled peptide showed good linearity in matrix solution. the content of ugt1a1 was 17.30 nmol/g protein and 18.23 nmol/g protein obtained by standard curve and stable isotope dilution method, respectively. in summary, the results obtained by two methods were consistent, however, the stable isotope dilution method was more convenient than by standard curve, and it was suitable for high throughput determination in complex system.
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	quantitative rt-pcr ; enzyme reactor ; proteins ; peptide ; safety
收录类别	SCI
语种	英语
WOS记录号	WOS:000333673500002
公开日期	2016-05-09
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/145619]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
作者单位	1.Jilin Univ, Coll Life Sci, Changchun 130012, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
推荐引用方式 GB/T 7714	Liu Xi-Dong,Zhu Jun,Cong Yu-Ting,et al. Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach[J]. chinese journal of analytical chemistry,2014,42(1):10-15.
APA	Liu Xi-Dong.,Zhu Jun.,Cong Yu-Ting.,Hu Liang-Hai.,Ye Ming-Liang.,...&Zou Han-Fa.(2014).Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach.chinese journal of analytical chemistry,42(1),10-15.
MLA	Liu Xi-Dong,et al."Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach".chinese journal of analytical chemistry 42.1(2014):10-15.