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新型人类细胞色素P450 2S1(CYP2S1)的鉴定及组织分布

Identification and tissue distribution of the novel human cytochrome P450 2S1 (CYP2S1).

作者信息

Rylander T, Neve E P, Ingelman-Sundberg M, Oscarson M

机构信息

Division of Molecular Toxicology, Institute of Environmental Medicine, Karolinska Institutet, SE-171 77 Stockholm, Sweden.

出版信息

Biochem Biophys Res Commun. 2001 Feb 23;281(2):529-35. doi: 10.1006/bbrc.2001.4390.

Abstract

With the aid of the htgs and dbEST databases, a novel cytochrome P450 cDNA was found by homology searches, and the corresponding gene was identified on chromosome 19. Nested PCR was used to amplify a full-length sequence of 1515 bp. The predicted 504 amino acid sequence displays 38--49% identity with CYP2 family members and the protein was designated CYP2S1. mRNA dot blot analysis demonstrated high expression levels in trachea, lung, stomach, small intestine, and spleen. The expression pattern was confirmed by Northern blot, which also revealed a single transcript of approximately 2.4 kb. Western blot analysis, using an antiserum directed against the C-terminus of the enzyme, detected a protein in human lung with the same mobility as recombinant CYP2S1. Subcellular fractionation and immunostaining revealed that CYP2S1 was localized in the endoplasmic reticulum. We conclude that CYP2S1 represents a novel abundantly expressed human P450 with potential importance for extrahepatic xenobiotic metabolism.

摘要

借助htgs和dbEST数据库,通过同源性搜索发现了一个新的细胞色素P450 cDNA,并在19号染色体上鉴定出了相应的基因。采用巢式PCR扩增出1515 bp的全长序列。预测的504个氨基酸序列与CYP2家族成员具有38%-49%的同一性,该蛋白被命名为CYP2S1。mRNA斑点印迹分析显示在气管、肺、胃、小肠和脾脏中表达水平较高。Northern印迹证实了这种表达模式,同时也揭示了一条约2.4 kb的单一转录本。使用针对该酶C末端的抗血清进行的蛋白质印迹分析,在人肺中检测到一种与重组CYP2S1迁移率相同的蛋白质。亚细胞分级分离和免疫染色显示CYP2S1定位于内质网。我们得出结论,CYP2S1代表一种新的高表达的人类P450,对肝外异源物质代谢具有潜在重要性。

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