Rotenone

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Rotenone

Description

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).

Abbreviation: Rot

Reference: Gnaiger 2014 MitoPathways, MiPNet09.12, MiPNet03.02

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)..


Preparation of 1 mM stock solution (dissolved in 99.9% ethanol)
  1. Weigh 0.39 mg of rotenone and dissolve in 1 ml of ethanol absolute (99.9%).
  2. Divide into 100 µl aliquots (use dark glass vials).
  3. 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.


Microalgae applications
For the specific case of microalgal cells it is recommended to prepare higher concentration:
Preparation of 5.5 mM stock solution (dissolved in 99.9% ethanol)
  1. Weigh 4.4 mg of rotenone and dissolve in 2 ml of ethanol absolute (99.9%).
  2. Divide into 200 µl aliquots (use dark glass vials).
  3. Store frozen at -20 °C.


O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume: 15 µL using a 25 µL syringe (2 mL O2k-Chamber).
  • Final concentration: 41 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






MitoPedia topics: Inhibitor