Molecular and functional characterization of UDP-glucuronosyltransferase 1A in cynomolgus macaques.
Molecular and functional characterization of UDP-glucuronosyltransferase 1A in cynomolgus macaques.
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食蟹猴 UDP-葡萄糖醛酸基转移酶 1A 的分子和功能表征。
DOI：
10.1016/j.bcp.2018.06.027
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Uno;Y.;Takahira;R.;Murayama;N.;Ishii;Y.;Ikenaka;Y.;Ishizuka;M.;Yamazaki;H.;and Ikushiro;S.
尿苷二磷酸葡萄糖醛酸转移酶（UGT）是内源性底物和外源性物质代谢所必需的药物代谢酶。UGT的分子特征已在人类中进行了广泛研究，但在食蟹猴（一种广泛用于药物代谢研究的非人灵长类动物）中仍有待研究。在本研究中，在食蟹猴中分离并表征了12种UGT 1A cDNA（UGT 1A 1、1A 2、1A 4A、1A 4 B、1A 5A、1A 5 B、1A 5C、1A 6、1A 7、1A 8、1A 9和1A 10）。由于无义突变，UGT 1A 5C cDNA不含完整的编码区，因此从进一步分析中排除。所有11种食蟹猴UGT 1A的氨基酸序列与人UGT 1A具有高度序列同一性（92-95%），并且在遗传学上与人UGT 1A接近。这些食蟹猴UGT 1A基因与人UGT 1A基因相似，具有共同的外显子2-5，并含有每个基因特有的可变外显子1。此外，食蟹猴和人UGT 1A基因簇位于基因组的相应区域。在分析的10种组织类型中，食蟹猴UGT 1A mRNA在肝脏、空肠和/或肾脏（药物代谢器官）中表达最丰富，与人UGT 1A相似。在这11种食蟹猴UGT 1A mRNA中，食蟹猴UGT 1A 2、UGT 1A 9和UGT 1A 10 mRNA分别在肝脏、肾脏和空肠中表达最丰富。食蟹猴肝微粒体和UGT 1A蛋白催化人UGT 1A催化的底物（包括4-甲基伞形酮、4-硝基苯酚、雌二醇、三氟拉嗪、5-羟色胺和丙泊酚）的葡萄糖醛酸化，但三氟拉嗪葡萄糖醛酸化不受任何食蟹猴UGT 1A蛋白的催化。这些结果表明，食蟹猴UGT 1As是与人UGT 1As具有分子相似性的功能酶。
UDP-glucuronosyltransferases (UGTs) are drug-metabolizing enzymes essential for the metabolism of endogenous substrates and xenobiotics. Molecular characteristics of UGTs have been extensively investigated in humans, but in cynomolgus macaques, a non-human primate species widely used in drug metabolism studies, remain to be investigated. In this study, 12 UGT1A cDNAs (UGT1A1, 1A2, 1A4A, 1A4B, 1A5A, 1A5B, 1A5C, 1A6, 1A7, 1A8, 1A9, and 1A10) were isolated and characterized in cynomolgus macaques. UGT1A5C cDNA did not contain a complete coding region due to nonsense mutations, and was excluded from further analysis. Amino acid sequences of all 11 cynomolgus UGT1As had high sequence identities (92–95%) with human UGT1As and were phylogenetically close to human UGT1As. These cynomolgusUGT1Agenes shared exons 2–5, and contained a variable exon 1 unique to each gene, similar to humanUGT1Agenes. Moreover, cynomolgus and humanUGT1Agene clusters were located in corresponding regions in the genome. Among the 10 tissue types analyzed, cynomolgus UGT1A mRNAs were most abundantly expressed in the liver, jejunum, and/or kidney, the drug-metabolizing organs, similar to human UGT1As. Among these 11 cynomolgus UGT1A mRNAs, cynomolgus UGT1A2, UGT1A9, and UGT1A10 mRNAs were most abundantly expressed in the liver, kidney, and jejunum, respectively. Cynomolgus liver microsomes and UGT1A proteins catalyzed glucuronidation of the substrates human UGT1As catalyze, including 4-methylumbelliferone, 4-nitrophenol, estradiol, trifluoperazine, serotonin, and propofol, although trifluoperazine glucuronidation was not catalyzed by any cynomolgus UGT1A proteins. These results suggest that cynomolgus UGT1As are functional enzymes with molecular similarities to human UGT1As.