Dimerization of human uridine diphosphate glucuronosyltransferase allozymes 1A1 and 1A9 alters their quercetin glucuronidation activities.
Dimerization of human uridine diphosphate glucuronosyltransferase allozymes 1A1 and 1A9 alters their quercetin glucuronidation activities.
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人尿苷二磷酸葡萄糖醛酸基转移酶同种酶 1A1 和 1A9 的二聚化改变了其槲皮素葡萄糖醛酸化活性
DOI：
10.1038/srep23763
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发表时间：
2016-03-30
影响因子：
4.6
通讯作者：
Zeng S
中科院分区：
文献类型：
--
作者：
Liu YQ;Yuan LM;Gao ZZ;Xiao YS;Sun HY;Yu LS;Zeng S
尿苷二磷酸葡萄糖醛酸转移酶1A（UGT 1A）是一个主要的II相药物代谢酶超家族，参与人体内内源性和外源性物质的葡萄糖醛酸化。据报道，UGT 1A基因的许多多态性抑制或降低UGT 1A活性。在这项研究中，两种UGT 1A 1等位酶，UGT 1A 1野生型和剪接突变体，以及UGT 1A 9野生型及其三种UGT 1A 9等位酶，UGT 1A 9 *2（C3 Y），UGT 1A 9 *3（M33 T）和UGT 1A 9 *5（D256 N）在Bac-to-Bac表达系统中进行单表达或双表达。通过荧光共振能量转移（FRET）和免疫共沉淀分析观察到UGT 1A 1或UGT 1A 9等位酶的二聚化。UGT 1A的SNPs改变了蛋白质-蛋白质相互作用的能力，导致不同的FRET效率和供体-受体距离。二聚化改变了UGT 1A 1和UGT 1A 9在槲皮素葡萄糖醛酸化中的化学区域选择性、底物结合亲和力和酶活性。这些发现为纯合和杂合UGT 1A 1和UGT 1A 9等位酶表达对槲皮素葡萄糖醛酸化的影响提供了分子见解。
Uridine diphosphate glucuronosyltransferase 1A (UGT1A) is a major phase II drug-metabolism enzyme superfamily involved in the glucuronidation of endobiotics and xenobiotics in humans. Many polymorphisms in UGT1A genes are reported to inhibit or decrease UGT1A activity. In this study, two UGT1A1 allozymes, UGT1A1 wild-type and a splice mutant, as well as UGT1A9 wild-type and its three UGT1A9 allozymes, UGT1A9*2(C3Y), UGT1A9*3(M33T) and UGT1A9*5(D256N) were single- or double-expressed in a Bac-to-Bac expression system. Dimerization of UGT1A1 or UGT1A9 allozymes was observed via fluorescence resonance energy transfer (FRET) and co-immunoprecipitation analysis. SNPs of UGT1A altered the ability of protein-protein interaction, resulting in differential FRET efficiencies and donor-acceptorrdistances. Dimerization changed the chemical regioselectivity, substrate-binding affinity and enzymatic activity of UGT1A1 and UGT1A9 in glucuronidation of quercetin. These findings provide molecular insights into the consequences of homozygous and heterozygous UGT1A1 and UGT1A9 allozymes expression on quercetin glucuronidation.