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Expression of CYP4F2 in human liver and kidney: assessment using specific peptide antibodies
Author(s) -
Lasker Jerome M.,
Hirani Vandana,
Yarovoy Anton,
Kozeska Anita,
Dhar Madhurima
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.921.5
Subject(s) - polyclonal antibodies , kidney , antibody , biochemistry , peptide , enzyme , chemistry , amino acid , biology , endocrinology , immunology
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 tissue‐specific expression and function of CYP4F2, one of four major CYP4F P450s found in human liver and kidney. Polyclonal peptide antibodies elicited in rabbits to CYP4F2 amino acid residues 61–74 (WGHQGMVNPTEEG) and 65–77 (GMVNPTEEGMRVL) recognized only CYP4F2 on immunoblots, and did not cross‐react with CYP4F3b, CYP4F11 or CYP4F12. Immunoquantitation with anti‐CYP4F2 peptide IgG showed highly‐variable CYP4F2 expression in liver (16.4 ± 18.6 pmol/mg; n = 28) and kidney cortex (3.9 ± 3.8 pmol/mg; n = 10), with two dissimilar subjects lacking the hepatic and renal enzyme entirely. Although CYP4F2 efficiently catalyzed ω‐hydroxylation of both arachidonate and oleate, only the former fatty acid‐metabolizing activity correlated strongly (r = 0.926; p < 0.025) with liver CYP4F2 content. Our study provides the first example of peptide antibodies that recognize a single CYP4F P450 found in liver and kidney, namely CYP4F2. Immunoquantitation and correlation analyses performed with these targeted antibodies indicate that CYP4F2 functions as a predominant arachidonate ω‐hydroxylase in human liver (supported by NIH AA07842).