Kinetic microplate-based assays for inhibitors of mitochondrial NADH:Ubiquinone oxidoreductase (complex I) and Succinate:Cytochrome c oxidoreductase

被引:14
作者
Jewess, PJ [1 ]
Devonshire, AL [1 ]
机构
[1] AFRC, Inst Arable Crops Res, Dept Biol & Ecol Chem, Harpenden AL5 2JQ, Herts, England
基金
英国生物技术与生命科学研究理事会;
关键词
96-well; assay; complex I; complex III; insect; microplate; mitochondria; NADH;
D O I
10.1006/abio.1999.4168
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 1999 Academic Press.
引用
收藏
页码:56 / 63
页数:8
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