Use of cloned and expressed human liver UDP-glucuronosyltransferases for analysis of drug glucuronide formation and assessment of drug toxicity

B. Burchell, T. Ebner, S. Baird, S. B. Senafi, D. Clarke, C. Brierley, L. Sutherland

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Five cloned human hepatic UDP-glucuronosyltransferase (UGT) cDNAs were stably expressed in tissue culture cell lines. More than 100 drugs and xenobiotics were used as substrates for glucuronidation catalyzed by the cloned human transferases to determine the chemical structures accepted as substrates. UGT-HP1 exhibited a limited substrate specificity for planar phenolic compounds, whereas UGT-HP4 was more accepting of nonplanar phenols, anthraquinones, flavones, alphatic alcohols, aromatic carboxylic acids, steroids and many drugs of varied structure. UGT-HP3 (bilirubin UGT) catalyzed the glucuronidation of ethinylestradiol. UGT-H6 and UGT-H25 (steroid bile acid UGTs) also catalyzed the glucuronidation of some xenobiotics. Levels of UGT-HP4 activity towards some substrates were sufficient to allow determination of kinetic parameters for the enzyme reaction. Further, metabolism of drugs could be studied by addition to the recombinant cell lines in culture and extraction of the media allowed analysis of glucuronide formation. The protection afforded against cytotoxic drugs was observed. The data presented here demonstrate the potential of using these recombinant cell lines for investigation of phase II metabolism by human UGTs and subtle differences in protein structure which affect their specificity.

LanguageEnglish
Pages19-23
Number of pages5
JournalEnvironmental Health Perspectives
Volume102
Issue numberSUPPL. 9
DOIs
Publication statusPublished - 1 Nov 1994
Externally publishedYes

Fingerprint

Glucuronosyltransferase
Glucuronides
Drug-Related Side Effects and Adverse Reactions
Liver
Pharmaceutical Preparations
bilirubin glucuronoside glucuronosyltransferase
Xenobiotics
Cell Line
Steroids
Flavones
Anthraquinones
Ethinyl Estradiol
Phenols
Transferases
Substrate Specificity
Carboxylic Acids
Bile Acids and Salts
Culture Media
Complementary DNA
Alcohols

Cite this

Burchell, B. ; Ebner, T. ; Baird, S. ; Senafi, S. B. ; Clarke, D. ; Brierley, C. ; Sutherland, L. / Use of cloned and expressed human liver UDP-glucuronosyltransferases for analysis of drug glucuronide formation and assessment of drug toxicity. In: Environmental Health Perspectives. 1994 ; Vol. 102, No. SUPPL. 9. pp. 19-23.
@article{a4552682de464a5499bb5ae8e470d460,
title = "Use of cloned and expressed human liver UDP-glucuronosyltransferases for analysis of drug glucuronide formation and assessment of drug toxicity",
abstract = "Five cloned human hepatic UDP-glucuronosyltransferase (UGT) cDNAs were stably expressed in tissue culture cell lines. More than 100 drugs and xenobiotics were used as substrates for glucuronidation catalyzed by the cloned human transferases to determine the chemical structures accepted as substrates. UGT-HP1 exhibited a limited substrate specificity for planar phenolic compounds, whereas UGT-HP4 was more accepting of nonplanar phenols, anthraquinones, flavones, alphatic alcohols, aromatic carboxylic acids, steroids and many drugs of varied structure. UGT-HP3 (bilirubin UGT) catalyzed the glucuronidation of ethinylestradiol. UGT-H6 and UGT-H25 (steroid bile acid UGTs) also catalyzed the glucuronidation of some xenobiotics. Levels of UGT-HP4 activity towards some substrates were sufficient to allow determination of kinetic parameters for the enzyme reaction. Further, metabolism of drugs could be studied by addition to the recombinant cell lines in culture and extraction of the media allowed analysis of glucuronide formation. The protection afforded against cytotoxic drugs was observed. The data presented here demonstrate the potential of using these recombinant cell lines for investigation of phase II metabolism by human UGTs and subtle differences in protein structure which affect their specificity.",
keywords = "Bilirubin, Multigene family, Phenols, Stable expression, Steroids",
author = "B. Burchell and T. Ebner and S. Baird and Senafi, {S. B.} and D. Clarke and C. Brierley and L. Sutherland",
year = "1994",
month = "11",
day = "1",
doi = "10.1289/ehp.94102s919",
language = "English",
volume = "102",
pages = "19--23",
journal = "Environmental Health Perspectives",
issn = "0091-6765",
publisher = "Public Health Services, US Dept of Health and Human Services",
number = "SUPPL. 9",

}

Use of cloned and expressed human liver UDP-glucuronosyltransferases for analysis of drug glucuronide formation and assessment of drug toxicity. / Burchell, B.; Ebner, T.; Baird, S.; Senafi, S. B.; Clarke, D.; Brierley, C.; Sutherland, L.

In: Environmental Health Perspectives, Vol. 102, No. SUPPL. 9, 01.11.1994, p. 19-23.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Use of cloned and expressed human liver UDP-glucuronosyltransferases for analysis of drug glucuronide formation and assessment of drug toxicity

AU - Burchell, B.

AU - Ebner, T.

AU - Baird, S.

AU - Senafi, S. B.

AU - Clarke, D.

AU - Brierley, C.

AU - Sutherland, L.

PY - 1994/11/1

Y1 - 1994/11/1

N2 - Five cloned human hepatic UDP-glucuronosyltransferase (UGT) cDNAs were stably expressed in tissue culture cell lines. More than 100 drugs and xenobiotics were used as substrates for glucuronidation catalyzed by the cloned human transferases to determine the chemical structures accepted as substrates. UGT-HP1 exhibited a limited substrate specificity for planar phenolic compounds, whereas UGT-HP4 was more accepting of nonplanar phenols, anthraquinones, flavones, alphatic alcohols, aromatic carboxylic acids, steroids and many drugs of varied structure. UGT-HP3 (bilirubin UGT) catalyzed the glucuronidation of ethinylestradiol. UGT-H6 and UGT-H25 (steroid bile acid UGTs) also catalyzed the glucuronidation of some xenobiotics. Levels of UGT-HP4 activity towards some substrates were sufficient to allow determination of kinetic parameters for the enzyme reaction. Further, metabolism of drugs could be studied by addition to the recombinant cell lines in culture and extraction of the media allowed analysis of glucuronide formation. The protection afforded against cytotoxic drugs was observed. The data presented here demonstrate the potential of using these recombinant cell lines for investigation of phase II metabolism by human UGTs and subtle differences in protein structure which affect their specificity.

AB - Five cloned human hepatic UDP-glucuronosyltransferase (UGT) cDNAs were stably expressed in tissue culture cell lines. More than 100 drugs and xenobiotics were used as substrates for glucuronidation catalyzed by the cloned human transferases to determine the chemical structures accepted as substrates. UGT-HP1 exhibited a limited substrate specificity for planar phenolic compounds, whereas UGT-HP4 was more accepting of nonplanar phenols, anthraquinones, flavones, alphatic alcohols, aromatic carboxylic acids, steroids and many drugs of varied structure. UGT-HP3 (bilirubin UGT) catalyzed the glucuronidation of ethinylestradiol. UGT-H6 and UGT-H25 (steroid bile acid UGTs) also catalyzed the glucuronidation of some xenobiotics. Levels of UGT-HP4 activity towards some substrates were sufficient to allow determination of kinetic parameters for the enzyme reaction. Further, metabolism of drugs could be studied by addition to the recombinant cell lines in culture and extraction of the media allowed analysis of glucuronide formation. The protection afforded against cytotoxic drugs was observed. The data presented here demonstrate the potential of using these recombinant cell lines for investigation of phase II metabolism by human UGTs and subtle differences in protein structure which affect their specificity.

KW - Bilirubin

KW - Multigene family

KW - Phenols

KW - Stable expression

KW - Steroids

UR - http://www.scopus.com/inward/record.url?scp=0028595707&partnerID=8YFLogxK

U2 - 10.1289/ehp.94102s919

DO - 10.1289/ehp.94102s919

M3 - Article

VL - 102

SP - 19

EP - 23

JO - Environmental Health Perspectives

T2 - Environmental Health Perspectives

JF - Environmental Health Perspectives

SN - 0091-6765

IS - SUPPL. 9

ER -