![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
S Fournel-Gigleux, L Sutherland, N Sabolovic, B Burchell and G Siest
Centre du Medicament, URA CNRS 597, Nancy, France.
Two human liver UDP-glucuronosyltransferase cDNA clones (HLUGP1 and HLUG25) were individually inserted into the eukaryotic expression vector pKCRH2. Each recombinant plasmid was cotransfected with a SFVneo vector, thereby allowing establishment of several V79 cell lines retaining the exogenous UDP-glucuronosyltransferase cDNA after selection with G418 (Geneticin). Southern blot analysis suggested that the cDNAs were integrated into the host cell genome. Northern blot and immunoblot analyses indicated that the cDNAs were correctly transcribed and translated for the production of functional enzymes. The established recombinant V79 cell lines stably expressed the UDP- glucuronosyltransferase activities towards 1-naphthol (HLUGP1) and hyodeoxycholic acid (HLUG25) at levels 10-20-fold higher than with transient expression, and in the range found in human liver. These high levels of expression of UDP-glucuronosyltransferase activity allowed the determination of apparent kinetic constants and substrate specificities of glucuronidation in the genetically engineered cell lines. HLUG25 cDNA encoded an isoform with restricted specificity towards the 6-OH group of the bile acid hyodeoxycholic acid. The other steroids, bile acids, endobiotics, and xenobiotics tested as substrates were glucuronidated in various samples of human liver microsomes, but not by this isoenzyme. This study, allowing the expression of individual UDP-glucuronosyltransferases in heterologous cells with no endogenous transferases, offered a unique solution for the characterization of UDP-glucuronosyltransferase functional heterogeneity.
This article has been cited by other articles:
|
|
 |
|
  Y. Tochigi, N. Yamashiki, S. Ohgiya, S. Ganaha, and H. Yokota ISOFORM-SPECIFIC EXPRESSION AND INDUCTION OF UDP-GLUCURONOSYLTRANSFERASE IN IMMUNOACTIVATED PERITONEAL MACROPHAGES OF THE RAT Drug Metab. Dispos., September 1, 2005; 33(9): 1391 - 1398. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. G. Staines, M. W. H. Coughtrie, and B. Burchell N-Glucuronidation of Carbamazepine in Human Tissues Is Mediated by UGT2B7 J. Pharmacol. Exp. Ther., December 1, 2004; 311(3): 1131 - 1137. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Cummings, B. T. Ethell, L. Jardine, G. Boyd, J. S. Macpherson, B. Burchell, J. F. Smyth, and D. I. Jodrell Glucuronidation as a Mechanism of Intrinsic Drug Resistance in Human Colon Cancer: Reversal of Resistance by Food Additives Cancer Res., December 1, 2003; 63(23): 8443 - 8450. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Taskinen, B. T. Ethell, P. Pihlavisto, A. M. Hood, B. Burchell, and M. W. H. Coughtrie CONJUGATION OF CATECHOLS BY RECOMBINANT HUMAN SULFOTRANSFERASES, UDP-GLUCURONOSYLTRANSFERASES, AND SOLUBLE CATECHOL O-METHYLTRANSFERASE: STRUCTURE-CONJUGATION RELATIONSHIPS AND PREDICTIVE MODELS Drug Metab. Dispos., September 1, 2003; 31(9): 1187 - 1197. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Krishnaswamy, S. X. Duan, L. L. von Moltke, D. J. Greenblatt, and M. H. Court Validation of Serotonin (5-Hydroxtryptamine) as an in Vitro Substrate Probe for Human UDP-Glucuronosyltransferase (UGT) 1A6 Drug Metab. Dispos., January 1, 2003; 31(1): 133 - 139. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. T. Ethell, S. Ekins, J. Wang, and B. Burchell Quantitative Structure Activity Relationships for the Glucuronidation of Simple Phenols by Expressed Human UGT1A6 and UGT1A9 Drug Metab. Dispos., June 1, 2002; 30(6): 734 - 738. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Antonio, J.-P. Grillasca, J. Taskinen, E. Elovaara, B. Burchell, M.-H. Piet, B. Ethell, M. Ouzzine, S. Fournel-Gigleux, and J. Magdalou Characterization of Catechol Glucuronidation in Rat Liver Drug Metab. Dispos., February 1, 2002; 30(2): 199 - 207. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Ouzzine, L. Antonio, B. Burchell, P. Netter, S. Fournel-Gigleux, and J. Magdalou Importance of Histidine Residues for the Function of the Human Liver UDP-Glucuronosyltransferase UGT1A6: Evidence for the Catalytic Role of Histidine 370 Mol. Pharmacol., April 13, 2001; 58(6): 1609 - 1615. [Abstract] [Full Text]
|
|
|
|
 |
|
 D. Turgeon, J.-S. Carrier, E. Levesque, D. W. Hum, and A. Belanger Relative Enzymatic Activity, Protein Stability, and Tissue Distribution of Human Steroid-Metabolizing UGT2B Subfamily Members Endocrinology, February 1, 2001; 142(2): 778 - 787. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. T. Ethell, K. Beaumont, D. J. Rance, and B. Burchell Use of Cloned and Expressed Human UDP-Glucuronosyltransferases for the Assessment of Human Drug Conjugation and Identification of Potential Drug Interactions Drug Metab. Dispos., January 1, 2001; 29(1): 48 - 53. [Abstract] [Full Text]
|
|
|
|
 |
|
 A. Radominska-Pandya, G. Chen, V. M. Samokyszyn, J. M. Little, W. E. Gall, G. Zawada, N. Terrier, J. Magdalou, and P. Czernik Application of photoaffinity labeling with [3H] all trans- and 9-cis-retinoic acids for characterization of cellular retinoic acid-binding proteins I and II Protein Sci., January 1, 2001; 10(1): 200 - 211. [Abstract] [Full Text]
|
|
|
|
|
|
U. Breyer-Pfaff, U. Mey, M. D. Green, and T. R. Tephly Comparative N-Glucuronidation Kinetics of Ketotifen and Amitriptyline by Expressed Human UDP-Glucuronosyltransferases and Liver Microsomes Drug Metab. Dispos., August 1, 2000; 28(8): 869 - 872. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. Ouzzine, J. Magdalou, B. Burchell, and S. Fournel-Gigleux An Internal Signal Sequence Mediates the Targeting and Retention of the Human UDP-Glucuronosyltransferase 1A6 to the Endoplasmic Reticulum J. Biol. Chem., October 29, 1999; 274(44): 31401 - 31409. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Furlan, S. Demirdjian, O. Bourdon, J. Magdalou, and A.-M. Taburet Glucuronidation of Drugs by Hepatic Microsomes Derived from Healthy and Cirrhotic Human Livers J. Pharmacol. Exp. Ther., May 1, 1999; 289(2): 1169 - 1175. [Abstract] [Full Text]
|
|
|
|
 |
|
 L McLaughlin, B Burchell, M Pritchard, C. Wolf, and T Friedberg Treatment of mammalian cells with the endoplasmic reticulum-proliferator compactin strongly induces recombinant and endogenous xenobiotic metabolizing enzymes and 3-hydroxy-3-methylglutaryl-CoA reductase in vitro J. Cell Sci., January 2, 1999; 112(4): 515 - 523. [Abstract] [PDF]
|
|
|
|
|
|
C. Senay, M. Ouzzine, E. Battaglia, D. Pless, V. Cano, B. Burchell, A. Radominska, J. Magdalou, and S. Fournel-Gigleux Arginine 52 and Histidine 54 Located in a Conserved Amino-Terminal Hydrophobic Region (LX2-R52-G-H54-X3-V-L) Are Important Amino Acids for the Functional and Structural Integrity of the Human Liver UDP-Glucuronosyltransferase UGT1*6 Mol. Pharmacol., March 1, 1997; 51(3): 406 - 413. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
