Rotenone is an inhibitor of Complex I (CI) and thus inhibits NADH oxidation. It inhibits the transfer of electrons from iron-sulfur centers in CI to ubiquinone via binding to the ubiquinone binding site of CI. See also Complex II-linked substrate state, S(Rot).
MitoPedia topics: Inhibitor
Application in HRR
- Rot: Rotenone (C23H22O6); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (≥ 95%).
- Caution: Very toxic and light sensitive (store solution in a dark glass vial). Use fresh aliquots for every experimental day!.
- Preparation of 1 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 0.39 mg of rotenone and dissolve in 1 ml of ethanol absolute (99.9%).
- Divide into 100 µl aliquots (use dark glass vials).
- Store frozen at -20 °C.
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber).
- Final concentration: 0.5 µM.
Inhibition of CI by rotenone
- Fato R, Bergamini C, Bortolus M, Maniero AL, Leoni S, Ohnishi T, Lenaz G (2009) Differential effects of mitochondrial Complex I inhibitors on production of reactive oxygen species. Biochim Biophys Acta 1787:384–92. >>PMC2724837