Complete cDNA sequence and cDNA-directed expression of CYP4A11, a fatty acid omega-hydroxylase expressed in human kidney.
A cDNA was isolated from a human kidney lambda gt10 library using the rat CYP4A3 cDNA as a probe. The cDNA-deduced amino acid sequence encoded a protein of 519 amino acids that was designated CYP4A11 (Nelson et al., 1993) and exhibited 76%, 72%, 80%, and 53% similarities to rat CYP4A1, rat CYP4A3, rabbit CYP4A6, and human CYP4B1, respectively. The deduced amino-terminal amino acid sequence of this cDNA agreed with the amino-terminal amino acid sequence of a major P450 protein purified from human renal microsomes. A second variant form of CYP4A11 cDNA, designated CYP4A11v, was isolated from the same library and had a deletion of a single adenine residue, thereby extending the reading frame and resulting in a protein of 591 amino acids. CYP4A11v is probably encoded by a rare allelic variant of CYP4A11, since no mutant alleles were uncovered in 15 normal individuals, as determined by a polymerase chain reaction (PCR) diagnostic test. Baculovirus-mediated cDNA expression of CYP4A11 yielded a P450 protein having a lambda max of 452 nm when reduced and complexed with carbon monoxide. The expressed enzyme efficiently catalyzed omega-hydroxylation of lauric acid. No detectable activity was uncovered toward arachidonic acid and prostaglandin E1. The cDNA-expressed variant, CYP4A11v, was found to be unstable and not to efficiently metabolize lauric acid, as assessed by both baculovirus and monkey kidney COS cell cDNA expression systems. These studies indicate that CYP4A11 is a major fatty acid-metabolizing P450 that is expressed in human kidney.
Imaoka
S
S
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, MD 20892.
Ogawa
H
H
Kimura
S
S
Gonzalez
F J
FJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D042926:Cytochrome P-450 CYP4A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D013057:Spectrum Analysis
D018076:DNA, Complementary; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D042926:Cytochrome P-450 CYP4A