Document Detail


Protein quantification of UDP-glucuronosyltransferases 1A1 and 2B7 in human liver microsomes by LC-MS/MS and correlation with glucuronidation activities.
MedLine Citation:
PMID:  22435749     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The aims of this study were to quantify absolute protein levels of uridine 5'-diphosphate-glucuronosyltransferases (UGTs) 1A1 and 2B7 in human liver microsomes (HLMs) and to investigate their correlation with marker activities. A quantification method for UGT1A1 and UGT2B7 in HLMs was developed. Unique tryptic peptides of UGT1A1 and UGT2B7 in tryptically digested HLMs were simultaneously quantified by liquid chromatography (LC) equipped with tandem mass spectrometry (MS) using corresponding stable isotope-labelled peptides as internal standards. Bovine serum albumin was used as a blank matrix for calibration curve samples. Our procedure had good digestion efficiency, sensitivity, calibration curve linearity, and reproducibility of digestion to quantification. In 16 individual HLMs, the protein levels of UGT1A1 and UGT2B7 ranged from 6.50 to 44.6 pmol/mg and 4.45 to 18.2 pmol/mg, respectively. Estradiol 3β-glucuronidation correlated strongly with the UGT1A1 level, indicating its high reliability as a reaction marker. Both morphine 3-O- and 6-O-glucuronidation significantly correlated with UGT2B7 level. However, the intercept of the linear regression clearly indicates that morphine glucuronidation was mediated by other UGT isoforms in addition to UGT2B7.
Authors:
Yuichiro Sato; Masanori Nagata; Akio Kawamura; Aiji Miyashita; Takashi Usui
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Publication Detail:
Type:  Journal Article     Date:  2012-03-21
Journal Detail:
Title:  Xenobiotica; the fate of foreign compounds in biological systems     Volume:  42     ISSN:  1366-5928     ISO Abbreviation:  Xenobiotica     Publication Date:  2012 Sep 
Date Detail:
Created Date:  2012-08-08     Completed Date:  2012-12-10     Revised Date:  2014-03-28    
Medline Journal Info:
Nlm Unique ID:  1306665     Medline TA:  Xenobiotica     Country:  England    
Other Details:
Languages:  eng     Pagination:  823-9     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Biological Markers / analysis
Blotting, Western
Chromatography, Liquid
Electrophoresis, Polyacrylamide Gel
Estradiol / analogs & derivatives,  metabolism
Glucuronosyltransferase / analysis*
Humans
Isotope Labeling
Microsomes, Liver / enzymology*
Tandem Mass Spectrometry
Chemical
Reg. No./Substance:
0/Biological Markers; 15270-30-1/estradiol-3-glucuronide; 4TI98Z838E/Estradiol; EC 2.4.1.-/UGT1A1 enzyme; EC 2.4.1.-/UGT2B7 protein, human; EC 2.4.1.17/Glucuronosyltransferase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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