Gonzalez F J, Schmid B J, Umeno M, Mcbride O W, Hardwick J P, Meyer U A, Gelboin H V, Idle J R
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, MD 20892.
DNA. 1988 Mar;7(2):79-86. doi: 10.1089/dna.1988.7.79.
P450PCN protein levels and nifedipine oxidase activities were quantitated in 12 human livers and were shown to be highly correlated. Antibody against rat P450PCN1 completely inhibited all nifedipine oxidase activity in three human liver samples. These results suggest that a human P450 related to rat P450PCN1 is the major form of P450 catalyzing nifedipine oxidation. The cDNA for a human P450, designated phPCN1, was isolated from a human liver lambda gt11 cDNA library, and sequenced completely. The deduced amino acid sequence is 77% similar to rat P450PCN1. By use of the adenovirus- and SV40-based expression vecotr p91023(B), the phP450PCN1 cDNA was expressed in COS cells and had high nifedipine oxidase activity, providing conclusive evidence that this P450 is the primary enzyme associated with metabolism and inactivation of this important drug. Using somatic cell hybrids, the P450PCN gene was localized to human chromosome 7.
在12个人类肝脏中对P450PCN蛋白水平和硝苯地平氧化酶活性进行了定量分析，结果显示二者高度相关。抗大鼠P450PCN1抗体完全抑制了三个人类肝脏样本中的所有硝苯地平氧化酶活性。这些结果表明，与大鼠P450PCN1相关的一种人类P450是催化硝苯地平氧化的主要P450形式。从人类肝脏λgt11 cDNA文库中分离出一种人类P450的cDNA，命名为phPCN1，并对其进行了全序列测定。推导的氨基酸序列与大鼠P450PCN1有77%的相似性。通过使用基于腺病毒和SV40的表达载体p91023(B)，phP450PCN1 cDNA在COS细胞中得以表达，并具有高硝苯地平氧化酶活性，这为该P450是与这种重要药物的代谢和失活相关的主要酶提供了确凿证据。利用体细胞杂种，将P450PCN基因定位到人类第7号染色体上。
