Characterization of Microsomal Glutathione S-Transferases MGST1, MGST2, and MGST3 in Cynomolgus Macaques
Characterization of Microsomal Glutathione S-Transferases MGST1, MGST2, and MGST3 in Cynomolgus Macaques
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DOI：
10.1124/dmd.113.052977
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发表时间：
2013-09-01
影响因子：
3.9
通讯作者：
Yamazaki, Hiroshi
中科院分区：
文献类型：
--
作者：
Uno, Yasuhiro;Murayama, Norie;Yamazaki, Hiroshi
谷胱甘肽S-转移酶（GST）家族包括细胞溶质、线粒体和微粒体GST，所有必需的酶都代谢广泛的内源性和外源性底物。在人类的微粒体GST（MGSTs）中，MGST 1，MGST 2和MGST 3参与解毒;然而，在食蟹猴（一种广泛用于药物代谢和毒性研究的重要灵长类动物）中，MGSTs尚未得到充分研究。在本研究中，从肝组织中分离食蟹猴MGST 2和MGST 3 cDNA，并与先前分离的食蟹猴MGST 1一起进行沿着表征。为了与食蟹猴cDNA进行比较，还从恒河猴（与食蟹猴密切相关）肝脏分离MGST 2和MGST 3 cDNA。食蟹猴MGST 2和MGST 3分别与人MGST 2和MGST 3高度同源（99%和98%），并且与恒河猴直系同源物的氨基酸序列几乎相同，并且通过系统发育分析它们与人MGST 2和MGST 3紧密聚类。对基因组数据的分析表明，MGST 1、MGST 2和MGST 3分别在猕猴和人类中具有相似的基因结构和基因组组织。因此，食蟹猴MGST与相应的人MGST具有分子相似性。食蟹猴MGST 2和MGST 3在肝脏、空肠和肾脏中表达，但水平低于MGST 1。GST活性测定与1-氯-2，4-二硝基苯和1，2-环氧-3-（p-硝基苯氧基）丙烷作为底物，使用异源表达的蛋白质在大肠杆菌。食蟹猴MGST 1、MGST 2和MGST 3与1-氯-2，4-二硝基苯和1，2-环氧-3-（对硝基苯氧基）丙烷结合，表明食蟹猴MGST 1、MGST 2和MGST 3是功能性酶。这些结果表明，这些功能性食蟹猴MGST酶和相应的人MGST在分子上相似。
The glutathione S-transferase (GST) family comprises cytosolic, mitochondrial, and microsomal GSTs, all essential enzymes that metabolize a wide range of endogenous and exogenous substrates. Among the microsomal GSTs (MGSTs) in humans, MGST1, MGST2, and MGST3 are involved in detoxification; however, MGSTs have not been fully investigated in cynomolgus macaque, an important primate species widely used in drug metabolism and toxicity studies. In the present study, cynomolgus MGST2 and MGST3 cDNAs were isolated from liver tissue and characterized along with previously isolated cynomolgus MGST1. For comparison with the cynomolgus cDNAs, MGST2 and MGST3 cDNAs were also isolated from rhesus macaque (closely related to cynomolgus macaque) liver. Cynomolgus MGST2 and MGST3, respectively, were highly identical (99 and 98%) to human MGST2 and MGST3 and nearly identical to the amino acid sequences of the rhesus orthologs, and they were closely clustered with human MGST2 and MGST3 by phylogenetic analysis. The analysis of genome data indicated that MGST1, MGST2, and MGST3, respectively, had similar gene structures and genomic organization in macaque and human. Therefore, cynomolgus MGSTs have molecular similarities to the corresponding human MGSTs. Cynomolgus MGST2 and MGST3 were expressed in liver, jejunum, and kidney, but at lower levels than MGST1. GST activities were measured with 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy) propane as substrates, using proteins heterologously expressed in Escherichia coli. Cynomolgus MGST1, MGST2, and MGST3 conjugated 1-chloro-2,4-dinitrobenzene and 1,2-epoxy-3-(p-nitrophenoxy) propane, indicating that cynomolgus MGST1, MGST2, and MGST3 are functional enzymes. These results suggest that these functional cynomolgus MGST enzymes and the corresponding human MGSTs are molecularly similar.