Gene expression of a novel cytochrome P450 of the CYP4F subfamily in human seminal vesicles.
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.
Department of Pharmaceutical Biosciences, Uppsala University, Sweden. Johan.Bylund@farmbio.uu.se
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J
Bylund
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N
Finnström
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EH
Oliw
D000595:Amino Acid Sequence; D001189:Aryl Hydrocarbon Hydroxylases; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005091:Exons / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007438:Introns / Q000235:genetics; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008297:Male; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012669:Seminal Vesicles / Q000201:enzymology* / Q000378:metabolism; D016415:Sequence Alignment
D017931:DNA Primers; D007527:Isoenzymes; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C120451:CYP4F8 protein, human; C026961:prostaglandin omega hydroxylases