叙利亚仓鼠细胞色素P450 2D新异构体CYP2D20、CYP2D27和CYP2D28的分子克隆与鉴定

Tomohiro Oka , Morio Fukuhara , Fusao Ushio , Kouichi Kurose
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引用次数: 1

摘要

我们在叙利亚仓鼠(Mesocricetus auratus)中克隆了三个新的细胞色素P450 (CYP) 2D cdna。每个克隆包含1500个核苷酸的开放阅读框,编码500个氨基酸的蛋白质。这些克隆的氨基酸序列与其他CYP2D成员的氨基酸序列具有较高的一致性,因此,这些克隆被命名为CYP2D20、CYP2D27和CYP2D28。Northern blot分析显示,CYP2D27 mRNA在肝脏中表达,而在肾脏、小肠和脑中不表达,而CYP2D20和CYP2D28 mRNA在这些组织中未检测到。肝脏中CYP2D27 mRNA的表达不存在性别差异,3-甲基胆蒽和苯巴比妥均未诱导CYP2D27 mRNA表达。我们鉴定了重组CYP2D27在COS-7细胞中表达的酶活性。CYP2D27蛋白具有特定于CYP2D亚家族的丁醛醇1′-羟化酶和碎片醌4-羟化酶活性。
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Molecular cloning and characterization of three novel cytochrome P450 2D isoforms, CYP2D20, CYP2D27, and CYP2D28 in the Syrian hamster (Mesocricetus auratus)

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.

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