Molecular characterization of functional UDP-glucuronosyltransferases 1A and 2B in common marmosets
Molecular characterization of functional UDP-glucuronosyltransferases 1A and 2B in common marmosets
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普通狨猴功能性 UDP-葡萄糖醛酸基转移酶 1A 和 2B 的分子特征
DOI：
10.1016/j.bcp.2019.113748
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发表时间：
2020
影响因子：
5.8
通讯作者：
Yamazaki Hiroshi
中科院分区：
文献类型：
--
作者：
Uno Yasuhiro;Uehara Shotaro;Inoue Takashi;Kawamura Shu;Murayama Norie;Nishikawa Miyu;Ikushiro Shinichi;Sasaki Erika;Yamazaki Hiroshi
UDP-葡萄糖醛酸基转移酶(UGTS)是一种重要的药物结合酶，可代谢多种内生和异种底物。UGT的分子特征在人类中已经得到了广泛的研究，但仍有待于在普通绒猴身上进行研究，绒猴是一种广泛用于药物代谢研究的非人类灵长类物种。本研究分离并鉴定了11个UGT cDNA(UGT1A1、1A3、1A4、1A6、1A7和1A9；UGT2B49、2B50、2B51、2B52和2B53)。绒猴UGT1A与人UGT1A的序列同源性较高(89-93%)，而绒猴UGT2B的序列同源性较低(82-86%)。绒猴UGT与人类UGT的亲缘关系很近。就像人类UGT1Asdo一样，绒毛UGT1Agene共享外显子2-5，并包含每个基因唯一的可变外显子1；相反，绒毛UGT2B基因包含6个独特的外显子。此外，绒猴和人类UGT1A和UGT2B基因簇位于各自基因组中的相应区域。在所测试的五种组织类型中，绒猴UGT mRNAs在肝脏、空肠和/或肾脏中表达最丰富，即与人类UGT一样，在对药物代谢起重要作用的组织中表达。在所研究的11个绒猴UGT mRNAs中，绒猴UGT1A9、1A4和1A6mRNAs分别在肝脏、小肠和肾脏中表达最丰富。绒猴肝微粒体和重组UGT1A蛋白催化与人类UGT1AS催化的相同底物的葡萄糖醛酸化反应，包括雌二醇、三氟拉嗪、4-甲基伞形酮、5-羟色胺、4-硝基苯酚和异丙酚。在目前的条件下，三氟拉嗪可被绒猴肝微粒体糖醛酸化，但不能被UGT1A亚型中的任何一种所检测。这些结果共同表明，功能性绒猴UGT具有与人类UGT大致相似的分子特征。
UDP-glucuronosyltransferases (UGTs) are essential drug-conjugation enzymes that metabolize a variety of endobiotic and xenobiotic substrates. The molecular characteristics of UGTs have been extensively investigated in humans, but remain to be investigated in common marmosets, a nonhuman primate species widely used in drug metabolism studies. In this study, 11 UGT cDNAs (UGT1A1, 1A3, 1A4, 1A6, 1A7, and 1A9; and UGT2B49, 2B50, 2B51, 2B52, and 2B53) were isolated and characterized in marmosets. Marmoset UGT1As had high sequence identities (89–93%) with human UGT1As, but the sequence identities of marmoset UGT2Bs were lower (82–86%). Marmoset UGTs were found to be phylogenetically close to human UGTs. Just as humanUGT1Asdo, marmosetUGT1Agenes shared exons 2–5 and contained a variable exon 1 unique to each gene; in contrast, marmosetUGT2Bgenes contained six unique exons. Moreover, marmoset and humanUGT1AandUGT2Bgene clusters were located in corresponding regions in their respective genomes. Among the five tissue types tested, marmoset UGT mRNAs were most abundantly expressed in liver, jejunum, and/or kidney, i.e., in tissues important for drug metabolism, just as human UGTs are. Among the 11 marmoset UGT mRNAs investigated, marmoset UGT1A9, 1A4, and 1A6 mRNAs were the most abundantly expressed in liver, small intestine, and kidney, respectively. Marmoset liver microsomes and recombinant UGT1A proteins catalyzed the glucuronidation of the same substrates that human UGT1As catalyze, including estradiol, trifluoperazine, 4-methylumbelliferone, serotonin, 4-nitrophenol, and propofol. Trifluoperazine was glucuronidated by marmoset liver microsomes, but not by any of the UGT1A isoforms examined under the present conditions. These results collectively suggest that functional marmoset UGTs have generally similar molecular characteristics to human UGTs.