Antibodies to a synthetic peptide that react specifically with a common surface region on two hydrocarbon-inducible isoenzymes of cytochrome P-450 in the rat.
An antibody that reacts with two hydrocarbon-inducible isoenzymes of rat cytochrome P-450 (c and d) in the rat was produced by immunising with a synthetic peptide, Leu-Ile-Ser-Lys-Phe-Gln-Lys-Leu-Met, which has the same primary structure as that of a region of both of these isoenzymes. There was no crossreactivity with hydrocarbon-inducible isoenzymes in liver microsomes from rabbit, mouse or in man. Nor was there any crossreactivity detected with liver microsomes from uninduced rats, or rats induced with phenobarbitone or isonicotinic acid hydrazide. This is consistent with the primary structure of these isoenzymes in the regions aligned with amino acids 174-182 (the immunising peptide) in rat isoenzyme c and demonstrates the ability to produce antibodies of defined specificity against isoenzymes of cytochrome P-450 by using synthetic peptide. As the antibody preparation is able to bind to isoenzymes c and d in their native conformations, either as partially purified enzymes, or in microsomes, it is suggested that this region is present on the surface of these cytochromes P-450.
Edwards
R J
RJ
Department of Clinical Pharmacology, Royal Postgraduate Medical School, London, U.K.
Singleton
A M
AM
Sesardic
D
D
Boobis
A R
AR
Davies
D S
DS
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D003429:Cross Reactions; D003577:Cytochrome P-450 Enzyme System / Q000276:immunology*; D004790:Enzyme Induction; D004797:Enzyme-Linked Immunosorbent Assay; D006433:Hemocyanins / Q000378:metabolism; D006838:Hydrocarbons / Q000494:pharmacology*; D007527:Isoenzymes / Q000276:immunology*; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D006838:Hydrocarbons; D007527:Isoenzymes; D010455:Peptides; D008748:Methylcholanthrene; D006433:Hemocyanins; D003577:Cytochrome P-450 Enzyme System; C032808:keyhole-limpet hemocyanin