Difference between revisions of "Rotenone"

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Rotenone

Description

Rotenone is an (synonym. 2R,6aS,12aS)-1,2,6,6a,12,12a-hexahydro-2-isopropenyl-8,9 dimethoxychromeno[3,4-b]furo(2,3-h)chromen-6-on) inhibitor of complex I via inhibition of 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 S_(Rot).

Abbreviation: Rot

Reference: Gnaiger 2012 MitoPathways, MiPNet09.12, MiPNet03.02

MitoPedia topics: Inhibitor

Application in HRR

Rot: Rotenone (Rotenone ≥ 95%, C₂₃H₂₂O₆); Sigma R 8875, 1 g, store at R.T.; FW = 394.42

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):

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.

Oxygraph-2k manual titrations: MiPNet09.12

Titration volume: 1 µl using a 10 µl syringe.
Final conc. in 2 ml O2k-Chamber: 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: 384–392.

@@ Line 18: / Line 18: @@
-'''Preparation of 1 mM stock solution''' (dissolved in 99.9% Ethanol):
+'''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%)
+::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)
+::2. Divide into 100 µl aliquots (use dark glass vials).
-::3) store frozen at -20 °C
+::3) Store frozen at -20 °C.
-'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
+'''Oxygraph-2k manual titrations:'''  [[MiPNet09.12]]
+::* Titration volume: 1 µl using a 10 µl syringe.
+::* Final conc. in 2 ml O2k-Chamber: 0.5 µM.
-::* Titration volume: 1 µl using a 10 µl syringe
-::* Final conc. in 2 ml O2k-chamber: 0.5 µM
 == Inhibition of CI by rotenone ==
 * [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2724837/ 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: 384–392.]