叙利亚仓鼠（金黄地鼠）中三种新型细胞色素P450 2D亚型CYP2D20、CYP2D27和CYP2D28的分子克隆与特性分析

Molecular cloning and characterization of three novel cytochrome P450 2D isoforms, CYP2D20, CYP2D27, and CYP2D28 in the Syrian hamster (Mesocricetus auratus).

作者信息

Oka T, Fukuhara M, Ushio F, Kurose K

机构信息

Department ol Pharmceutical Sciences, National Institute of Public Health, Tokyo, Japan.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2000 Sep;127(2):143-52. doi: 10.1016/s0742-8413(00)00142-0.

DOI:10.1016/s0742-8413(00)00142-0

PMID:11083025

Abstract

We cloned three novel cytochrome P450 (CYP) 2D cDNAs in the Syrian hamster (Mesocricetus auratus). Each clone contained an open reading frame of 1500 nucleotides encoding a protein of 500 amino acids. The deduced amino acid sequences of these had high identities with those of the other CYP2D members, therefore, the clones were assigned as CYP2D20, CYP2D27, and CYP2D28. Northern blot analysis showed that the CYP2D27 mRNA was expressed in liver, but not in kidney, small intestine, and brain, while the CYP2D20 and CYP2D28 mRNAs were not detected in these tissues examined. The expression of CYP2D27 mRNA in liver did not show sex difference and was not induced by either 3-methylcholanthrene or phenobarbital treatment. We characterized the enzyme activities of recombinant CYP2D27 expressed in COS-7 cells. The CYP2D27 protein had the bufuralol 1'-hydroxylase and debrisoquine 4-hydroxylase activities that are specific to the CYP2D subfamily.

摘要

我们在叙利亚仓鼠（金仓鼠）中克隆了三个新的细胞色素P450（CYP）2D cDNA。每个克隆都包含一个1500个核苷酸的开放阅读框，编码一个由500个氨基酸组成的蛋白质。这些推导的氨基酸序列与其他CYP2D成员的序列具有高度同源性，因此，这些克隆被命名为CYP2D20、CYP2D27和CYP2D28。Northern印迹分析表明，CYP2D27 mRNA在肝脏中表达，但在肾脏、小肠和大脑中不表达，而在这些检测的组织中未检测到CYP2D20和CYP2D28 mRNA。CYP2D27 mRNA在肝脏中的表达没有性别差异，也不受3-甲基胆蒽或苯巴比妥处理的诱导。我们对在COS-7细胞中表达的重组CYP2D27的酶活性进行了表征。CYP2D27蛋白具有CYP2D亚家族特有的布非洛尔1'-羟化酶和异喹胍4-羟化酶活性。