Bylund J, Finnström N, Oliw E H
Department of Pharmaceutical Biosciences, Uppsala University, Sweden.
Biochem Biophys Res Commun. 1999 Jul 22;261(1):169-74. doi: 10.1006/bbrc.1999.1011.
19R-Hydroxyprostaglandins are major components of human seminal fluid. They are apparently formed in the seminal vesicles by NADPH-dependent omega2-hydroxylation. The hydroxylase is likely a cytochrome P450 (CYP), which has not been identified. To address this issue we studied gene expression of CYPs in human seminal vesicles (n = 4) with reverse-transcription polymerase chain reaction (RT-PCR). CYP1B1, CYP2E1, CYP2J2, CYP3A5, CYP4B1, and CYP4B1 with insertion of three nucleotides (Ser207) were detected in all subjects. RT-PCR with degenerate primers for the CYP4 family yielded a novel cDNA sequence, which was derived from a previously reported genomic sequence on chromosome 19p13.1 and present in all subjects. cDNA cloning showed that the deduced amino acid sequence consisted of 520 amino acids. Northern blot analysis demonstrated mRNA transcripts of approximately 2.1 and approximately 2.3 kb. The deduced protein showed 81.2 and 76.7% amino acid identity with the human enzymes CYP4F2 and CYP4F3. The novel CYP was designated CYP4F8.
19R-羟基前列腺素是人类精液的主要成分。它们显然是在精囊中由依赖NADPH的ω2-羟化作用形成的。这种羟化酶可能是一种细胞色素P450(CYP),但尚未被鉴定出来。为了解决这个问题,我们用逆转录聚合酶链反应(RT-PCR)研究了人类精囊(n = 4)中CYPs的基因表达。在所有受试者中均检测到了CYP1B1、CYP2E1、CYP2J2、CYP3A5、CYP4B1以及插入了三个核苷酸(Ser207)的CYP4B1。用针对CYP4家族的简并引物进行RT-PCR产生了一个新的cDNA序列,该序列源自先前报道的位于19号染色体p13.1上的基因组序列,且在所有受试者中均存在。cDNA克隆显示推导的氨基酸序列由520个氨基酸组成。Northern印迹分析显示有大约2.1 kb和大约2.3 kb的mRNA转录本。推导的蛋白质与人类酶CYP4F2和CYP4F3的氨基酸同一性分别为81.2%和76.7%。这种新的CYP被命名为CYP4F8。