Variability of human hepatic UDP-glucuronosyltransferase activity

被引:11
作者
Little, JM
Lester, R
Kuipers, F
Vonk, R
Mackenzie, PI
Drake, RR
Frame, L
Radominska-Pandya, A
机构
[1] Univ Arkansas Med Sci, Dept Biochem & Mol Biol, Little Rock, AR 72205 USA
[2] Univ Arkansas Med Sci, Dept Internal Med, Little Rock, AR 72205 USA
[3] Acad Hosp Groningen, Ctr Liver Digest & Metab Dis, Groningen, Netherlands
[4] Univ Groningen Hosp, Lab Nutr & Metab, Groningen, Netherlands
[5] Flinders Med Ctr, Dept Clin Pharmacol, Bedford Pk, SA, Australia
[6] Texas Tech Univ, Inst Environm & Human Hlth, Lubbock, TX 79409 USA
关键词
UDP-glucuronosyltransferase; human; liver; tyrosinemia;
D O I
10.18388/abp.1999_4168
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The availability of a unique series of liver samples from human subjects, both control patients (9) and those with liver disease (6; biliary atresia (2), retransplant, chronic tyrosinemia type I, tyrosinemia, Wilson's disease) allowed us to characterize human hepatic UDP-glucuronosyltransferases using photoaffinity labeling, immunoblotting and enzymatic assays. There was wide inter-individual variation in photoincorporation of the photoaffinity analogs, [P-32]5-azido-UDP-glucuronic acid and [P-32]5-azido-UDP-glucose and enzymatic glucuronidation of substrates specific to the two subfamilies of UDP-glucuronosyltransferases. However, the largest differences were between subjects with liver disease. Glucuronidation activities toward one substrate from each of the UDP-glucuronosyltransferases subfamilies, 1A and 2B, for control and liver disease, respectively, were 1.7-4.5 vs 0.4-4.7 nmol/mg x min for hyodeoxycholic acid (2B substrate) and 9.2-27.9 vs 8.1-75 nmol/mg x min for p-chloro-m-xylenol (1A substrate). Microsomes from a patient with chronic tyrosinemia (HL32) photoincorporated [P-32]5-azido-UDP-glucuronic acid at a level 1.5 times higher than the other samples, was intensely photolabeled by [P-32]5-azido-UDP-glucose and had significantly higher enzymatic activity toward p-chloro-m-xylenol. Immunoblot analysis using anti-UDP-glucuronosyltransferase antibodies demonstrated wide inter-individual variations in UDP-glucuronosyltransferase protein with increased UDP-glucuronosyltransferase protein in HL32 microsomes, corresponding to one of the bands photolabeled by both probes. Detailed investigation of substrate specificity, using substrates representative of both the 1A (bilirubin, 4-nitrophenol) and 2B (androsterone, testosterone) families was carried out with HL32, HL38 (age and sex matched control) and HL18 (older control). Strikingly increased (5-8-fold) glucuronidation activity was seen in comparison to HL18 only with the phenolic substrates. The results indicate that one or more phenol-specific UDP-glucuronosyltransferase 1A isoforms are expressed at above normal levels in this tyrosinemic subject.
引用
收藏
页码:351 / 363
页数:13
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