Monoclonal antibodies that inhibit enzyme activity of 3-methylcholanthrene-induced cytochrome P-450.

作者：Park Ss, T Fujino, Donna West, Frederick Peter Guengerich, Gelboin Hv · 发表于：PubMed · 年份：1982 · 被引用次数：168 · 研究领域：Monoclonal and Polyclonal Antibodies Research、Synthesis and Biological Evaluation、Pharmacogenetics and Drug Metabolism

Abstract Somatic cell hybrids were made between mouse myeloma cells and spleen cells derived from BALB/c mice immunized with liver microsomal cytochrome P-450 purified from rats treated with 3-methylcholanthrene (MC-P-450). Thirty-seven independent hybrid clones among 66 tested produced monoclonal antibodies to the MC-P-450 as measured by radioimmunoassay. More than 10 of the monoclonal antibodies formed were positive for MC-P-450 with respect to protein binding measured by radioimmunoassay, precipitation of the enzyme caused by antibody binding and enzyme aggregation, and inhibition of enzymatic activity. Analysis by gel electrophoresis indicated that a single microsomal protein band interacted with the antibody and that this band comigrated with MC-P-450. These monoclonal antibodies interacted with the major form of cytochrome P-450 from β-naphthoflavone-induced rats as well as with MC-P-450 but did not bind, precipitate, or inhibit the activity of the major form of cytochrome P-450 from phenobarbital-treated rats. The monoclonal antibodies inhibited 7-ethoxycoumarin deethylase and benzo( a )pyrene hydroxylation activity of the purified MC-P-450 with varying degrees, up to 90%, the latter as measured by the aryl hydrocarbon hydroxylase assay for phenol production. Analysis of benzo( a )pyrene metabolism by high-pressure liquid chromatography indicated that the monoclonal antibodies inhibited the enzyme activity of the purified MC-P-450 at all of the positions at which oxida...