STRUCTURE-FUNCTION OF HUMAN UDP-GLUCURONOSYLTRANSFERASES

描述（改编自申请人摘要）：葡萄糖醛酸化是一种 是外源性微生物生物转化和排泄的重要途径 药物和其它外源性物质和内源性化合物，例如胆红素， 类固醇激素、胆汁和脂肪酸以及甲状腺激素。 与葡萄糖醛酸缀合通常导致葡萄糖醛酸的增加。 溶解性和降低的化合物生物活性。然而，在这方面， 葡萄糖醛酸化可能参与化合物的生物活化， 诱变剂、致癌剂或其他药理活性形式。UDP- 葡萄糖醛酸基转移酶（UGT）是一个负责 葡萄糖醛酸化。本提案侧重于表达 人UGT cDNA在合适的宿主细胞中的表达以及UGT cDNA的表征 在不存在干扰UGT的情况下克隆蛋白的结构 同工酶主要目标之一是阐明 人UGT的多底物特异性在其初级 和三级结构，以了解葡萄糖醛酸化的作用 在内源性底物、药物和外源性物质的代谢中 人类该项目将在两个实验室（大学）进行。 阿肯色州大学和法国南希大学）， 在人类重组UGT研究方面的专业知识。这些研究将 适用于以下四种重组人UGT同工酶; 生理上重要的UGT，UGT 2B 4（猪去氧胆酸特异性， UGT 1 *1胆红素特异性），以及UGT*6（小，平面酚- 特异性和UGT 2B 10，一种未知底物特异性的新型UGT。 有两个互补的目标：（1）合成人重组 UGT的量足以表征、纯化、产生抗体 （2）研究人体结构 重组UGT及其与功能特性的关系 个体异构体。

DESCRIPTION (adapted from applicant's abstract): Glucuronidation is an important pathway in the biotransformation and excretion of exogenous drugs and other xenobiotics and endogenous compounds, such as bilirubin, steroid hormones, bile and fatty acids, and thyroid hormones. Conjugation with glucuronic acid generally results in the increased solubility and decreased biological activity of a compound. However, glucuronidation may be involved in the bioactivation of compounds to mutagens, carcinogens or other pharmacological active forms. UDP- glucuronosyltransferases (UGTs) are a family of enzymes responsible for glucuronidation. The present proposal focuses on the expression of human UGT cDNA in suitable host cells and characterization of the structure of the cloned protein in the absence of interfering UGTs isoenzyme. One of the principal goals will be to elucidate how the multi-substrate specificity of human UGTs are encoded in their primary and tertiary structure in order to understand the role of glucuronidation in the metabolism of endogenous substrates, drugs, and xenobiotics in humans. The project will be carried out in two laboratories (University of Arkansas and University of Nancy, France) with complementary expertise in the study of human recombinant UGTs. The studies will be applied to the following four recombinant human UGT isoenzyme; two physiologically important UGTs, UGT2B4 (hyodeoxycholic acid-specific and UGT1*1 bilirubin specific), as well as UGT*6 (small, planar phenol- specific and UGT2B10, a novel UGT of unknown substrate specificity. There are two complementary goals: (1) to synthesize human recombinant UGTs in amounts sufficient to characterize, purify, produce antibodies nd for structural studies; (2) to study the structure of human recombinant UGTs and their relationship to the functional properties of the individual isoforms.