Difference between revisions of "Rotenone"

Revision as of 18:06, 17 July 2016

high-resolution terminology - matching measurements at high-resolution

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

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

@@ Line 13: / Line 13: @@
 == Application in [[HRR]] ==
-'''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (≥ 95%).
+:::: '''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (≥ 95%).
-<span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial). Use fresh aliquots for every experimental day!</span>.
+:::: <span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial). Use fresh aliquots for every experimental day!</span>.
-'''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%).
+::::# 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).
+::::# Divide into 100 µl aliquots (use dark glass vials).
-::3) Store frozen at -20 °C.
+::::# Store frozen at -20 °C.
-'''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
+::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
-::* Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber).
+::::* Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber).
-::* Final concentration: 0.5 µM.
+::::* 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: 384–392. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2724837/ >>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 1787:384–92. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2724837/ >>PMC2724837]