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Absolute Quantification of Cytochrome P450 and Uridinediphosphate Glucuronosyl Transferase Isoforms by Proteomics-based Approach
Liu Xi-Dong1; Zhu Jun2; Cong Yu-Ting1; Hu Liang-Hai1; Ye Ming-Liang2; Gu Jing-Kai1; Zou Han-Fa2
KeywordUridinediphosphate Glucuronosyl Transferase Multiple Reaction Monitoring Liver Microsome Cytochrome P450 Liquid Chromatography-tandem Mass Spectrometry
Source PublicationCHINESE JOURNAL OF ANALYTICAL CHEMISTRY
2014
DOI10.3724/SP.J.1096.2014.30741
Volume42Issue:1Pages:10-15
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordQUANTITATIVE RT-PCR ; ENZYME REACTOR ; PROTEINS ; PEPTIDE ; SAFETY
AbstractStrategy 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.
Language英语
WOS IDWOS:000333673500002
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/145619
Collection中国科学院大连化学物理研究所
Affiliation1.Jilin Univ, Coll Life Sci, Changchun 130012, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
Recommended Citation
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.
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