Epidermal cytochrome P-450: immunochemical characterization of isoform induced by topical application of 3-methylcholanthrene to neonatal rat.
To further define the capacity of skin to metabolize various drugs and carcinogens by the cytochrome P-450 (P-450)-dependent monooxygenase, we studied the profile of P-450 isoform(s) in neonatal rat epidermis after application of 3-methylcholanthrene (3-MC) to skin. Highly specific monoclonal antibodies (MAbs) 2-66-3 and 1-7-1 against purified rat liver P-450 isoforms b and c induced by phenobarbital and 3-MC, respectively, were used. After a single topical application of 3-MC to rats aryl hydrocarbon hydroxylase (AHH), 7-ethoxycoumarin-O-deethylase and 7-ethoxyresorufin-O-deethylase activities in epidermal microsomes were induced significantly. Radioimmunoassay of epidermal microsomes from untreated control animals with 35S-labeled MAb-2-66-3 showed significant binding whereas negligible binding with MAb 1-7-1 was observed. On the other hand significant binding was observed with 35S-labeled MAb 1-7-1 with epidermal microsomes prepared from 3-MC-treated animals. Histochemical staining of epidermis from control animals showed no immunoreactivity with either MAb 1-7-1 or MAb 2-66-3 whereas epidermis obtained from 3-MC-treated rats showed significant immunoreactivity with MAb 1-7-1. Western blot analysis of epidermal microsomes prepared from control animals showed no immunoreactivity with either MAb 1-7-1 or 2-66-3 whereas epidermal microsomes prepared from 3-MC-treated animals showed distinct immunoreactivity with MAb 1-7-1. MAb 2-66-3 inhibited AHH and 7-ethoxycoumarin-O-deethylase activity (40-50%) in microsomes prepared from control animals.(ABSTRACT TRUNCATED AT 250 WORDS)
Khan
W A
WA
Department of Dermatology, Case Western Reserve University, Cleveland, Ohio.
Park
S S
SS
Gelboin
H V
HV
Bickers
D R
DR
Mukhtar
H
H
D000279:Administration, Cutaneous; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004817:Epidermis / Q000187:drug effects / Q000201:enzymology*; D066298:In Vitro Techniques; D008748:Methylcholanthrene / Q000494:pharmacology*; D010105:Oxygenases / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases