Kinetic microplate-based assays for inhibitors of mitochondrial NADH:ubiquinone oxidoreductase (complex I) and succinate:cytochrome c oxidoreductase.
Kinetic microplate-based assays for both mitochondrial NADH:ubiquinone oxidoreductase (complex I) and succinate:cytochrome c oxidoreductase using insect submitochondrial particles as the source of the enzyme activities have been developed. These assays have been used to design high-throughput screens for inhibitors of these mitochondrial electron transfer activities to assess their intrinsic in vitro efficacies as potential pesticides. These methods can be used to test up to 60 compounds per day without the use of automated sample handling and diluting technology. The accuracy, specificity, and reproducibility of the microplate methods compared well with conventional spectrophotometer-based assays.
Department of Biological and Ecological Chemistry, IACR-Rothamsted, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom.
P J
PJ
Jewess
A L
AL
Devonshire
D000818:Animals; D004175:Diptera / Q000201:enzymology; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000032:analysis*; D005069:Evaluation Studies as Topic; D066298:In Vitro Techniques; D007700:Kinetics; D008931:Mitochondria, Muscle / Q000201:enzymology; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors*; D015203:Reproducibility of Results; D013053:Spectrophotometry; D013367:Submitochondrial Particles / Q000201:enzymology; D013384:Succinate Cytochrome c Oxidoreductase / Q000037:antagonists & inhibitors*
D004791:Enzyme Inhibitors; D013384:Succinate Cytochrome c Oxidoreductase; D016660:NAD(P)H Dehydrogenase (Quinone)