Methoxyresorufin as a substrate for the fluorometric assay of insect microsomal O-dealkylases |
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| Authors: | Richard T Mayer John W Jermyn MDanny Burke Russell A Prough |
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| Institution: | 1. Veterinary Toxicology and Entomology Research Laboratory, Agricultural Research Service, U. S. Department of Agriculture, College Station, Texas 77840 USA;2. Biochemistry Department, Texas A&M University, College Station, Texas 77843 USA;3. Biochemistry Department, University of Texas Health Science Center, Dallas, Texas 75235 USA |
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| Abstract: | Methoxy-, ethoxy-, propoxy-, and butoxyresorufin were prepared and tested as substrates for the fluorometric assay of O-dealkylation by the mixed-function oxidase system in house flies, Musca domestica L. Methoxyresorufin proved to be the most suitable substrate because of the favorable reaction rates. This sensitive assay can be performed with a minimum of microsomal protein (<0.3 mg/ml) in 5 min or less. The apparent Km and maximum velocity were calculated as 2.88 μM and 0.27 nmol of resorufin produced/min/mg of microsomal protein, respectively. The O-dealkylation reaction required O2 and NADPH and was inhibited by CO. |
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