Isolation and sequence determination of a cDNA clone related to human cytochrome P-450 nifedipine oxidase.
Isolation and sequence determination of a cDNA clone related to human cytochrome P-450 nifedipine oxidase.
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与人细胞色素 P-450 硝苯地平氧化酶相关的 cDNA 克隆的分离和序列测定。
DOI：
10.1073/pnas.83.21.8064
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发表时间：
1986
影响因子：
11.1
通讯作者：
Guengerich,FP
中科院分区：
文献类型：
--
作者：
Beaune,PH;Umbenhauer,DR;Bork,RW;Lloyd,RS;Guengerich,FP
人肝细胞色素P-450 NF是负责钙通道阻滞剂硝苯地平氧化的细胞色素P-450的形式，据报道，硝苯地平在临床研究中显示出多态性。通过筛选λ gt 11噬菌体表达cDNA文库，我们分离到两个克隆：插入长度为0.8个碱基的NF 95和插入长度为2.2个碱基的NF 25。对两个克隆进行测序，并显示其重叠部分相同。NF 25的序列与大鼠细胞色素“P-450 PCN”cDNA（PCN =异烯醇酮-16 α-甲腈）的报道序列有77%相似。当该序列与属于其他家族的人细胞色素P-450比较时，相似性降低到45-53%[即，“pH P-450（1）”、“P1-450”、“P3-450”和“P-450MP”。推导的氨基酸序列与大鼠细胞色素P-450 PCN的相似性为73%，前21个氨基酸与人肝细胞色素P-450 p的报道相同。这些克隆的切片被切口翻译，并用作分析人类mRNA和基因组DNA的探针。条带的数量和大小表明P-450 NF属于多基因家族，即所谓的双烯醇酮-16 α-腈诱导型家族。
Human liver cytochrome P-450NF is the form of cytochrome P-450 responsible for the oxidation of the calcium-channel blocker nifedipine, which has been reported to show polymorphism in clinical studies. By screening a bacteriophage lambda gt11 expression cDNA library, we isolated two clones: NF95 with an insert length of 0.8 kilobases which gave a stable fusion protein and NF25 with an insert length of 2.2 kilobases. The two clones were both sequenced and shown to be identical in their overlapping section. The sequence of NF25 is 77% similar to that reported for a rat cytochrome "P-450PCN" cDNA (PCN = pregnenolone-16 alpha-carbonitrile). The similarity decreases to 45-53% when the sequence is compared to human cytochromes P-450 belonging to other families [i.e., "pH P-450(1)," "P1-450," "P3-450," and "P-450MP." The deduced amino acid sequence is 73% similar to that of rat cytochrome P-450PCN, and the first 21 amino acids are identical to those reported for human liver cytochrome "P-450p." Sections of these clones were nick-translated and used as probes for analyses of human mRNA and genomic DNA. The number and size of bands indicate that P-450NF belongs to a multigene family, the so-called pregnenolone-16 alpha-carbonitrile-inducible family.