Expression of CYP4F2 in human liver and kidney: assessment using targeted peptide antibodies.
P450 enzymes comprising the human CYP4F gene subfamily are catalysts of eicosanoid (e.g., 20-HETE and leukotriene B4) formation and degradation, although the role that individual CYP4F proteins play in these metabolic processes is not well defined. Thus, we developed antibodies to assess the tissue-specific expression and function of CYP4F2, one of four CYP4F P450s found in human liver and kidney. Peptide antibodies elicited in rabbits to CYP4F2 amino acid residues 61-74 (WGHQGMVNPTEEG) and 65-77 (GMVNPTEEGMRVL) recognized on immunoblots only CYP4F2 and not CYP4F3b, CYP4F11 or CYP4F12. Immunoquantitation with anti-CYP4F2 peptide IgG showed highly variable CYP4F2 expression in liver (16.4+/-18.6pmol/mg microsomal protein; n=29) and kidney cortex (3.9+/-3.8 pmol/mg; n=10), with two subjects lacking the hepatic or renal enzyme entirely. CYP4F2 content in liver microsomes was significantly correlated (r> or =0.63; p<0.05) with leukotriene B4 and arachidonate omega-hydroxylase activities, which are both CYP4F2-catalyzed. Our study provides the first example of a peptide antibody that recognizes a single CYP4F P450 expressed in human liver and kidney, namely CYP4F2. Immunoquantitation and correlation analyses performed with this antibody suggest that CYP4F2 functions as a predominant LTB4 and arachidonate omega-hydroxylase in human liver.
Hirani
Vandana
V
Jurist Institute for Biomedical Research, Hackensack University Medical Center, 30 Prospect Avenue, Hackensack, NJ 07601, USA.
Yarovoy
Anton
A
Kozeska
Anita
A
Magnusson
Ronald P
RP
Lasker
Jerome M
JM
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry*; D002648:Child; D002675:Child, Preschool; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis*; D000072470:Cytochrome P450 Family 4; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007223:Infant; D007668:Kidney / Q000201:enzymology*; D008099:Liver / Q000201:enzymology* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D000906:Antibodies; D003577:Cytochrome P-450 Enzyme System; D000072470:Cytochrome P450 Family 4; C117480:CYP4F2 protein, human