Difference between revisions of "Succinate"
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::: '''Preparation of 1 M stock solution''' (dissolved in H<sub>2</sub>O) | ::: '''Preparation of 1 M stock solution''' (dissolved in H<sub>2</sub>O) | ||
::::# | ::::# Weight 1.3505 g of succinate and dissolve in 3 mL H<sub>2</sub>O; | ||
::::# Check pH and adjust to 7.0 if necessary with 1 M HCl (usually the pH is 7 without any adjustment) | ::::# Check pH and adjust to 7.0 if necessary with 1 M HCl (usually the pH is 7 without any adjustment); | ||
::::# Transfer to 5 | ::::# Transfer to 5 mL volumetric glass flask and adjust the final volume to 5 mL; | ||
::::# Divide into 0.5 | ::::# Divide into 0.5 mL portions; | ||
::::# Store frozen at -20 °C. | ::::# Store frozen at -20 °C. | ||
Line 24: | Line 24: | ||
::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::::* In the absence of CI-linked substrates, add the CI-inhibitor [[rotenone]] before addition of succinate, to avoid accumulation of [[oxaloacetate]] with subsequent inhibition of | ::::* In the absence of CI-linked substrates, add the CI-inhibitor [[rotenone]] before addition of succinate, to avoid accumulation of [[oxaloacetate]] with subsequent inhibition of [[succinate dehydrogenase]]. See: [[Succinate and rotenone]]. | ||
::::* When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm the stock solution in your hands if necessary. | ::::* When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm the stock solution in your hands if necessary. | ||
::::* Titration volume: 20 | ::::* Titration volume: 20 µL using a 50 µL syringe (2 mL O2k-chamber). | ||
::::* Final concentration: 10 mM. | ::::* Final concentration: 10 mM. |
Revision as of 12:01, 14 August 2018
Description
Succinic acid, C4H6O4, (butanedioic acid) is a dicarboxylic acid which occurs under physiological conditions as the anion succinate2-, S, with pKa1 = 4.2 and pKa2 = 5.6. Succinate is formed in the TCA cycle, and is a substrate of CII, reacting to fumarate and feeding electrons into the Q-junction. Succinate (CII-linked) and NADH (CI-linked) provide convergent electron entries into the Q-junction. Succinate is transported across the inner mt-membrane by the dicarboxylate carrier. The plasma membrane of many cell types is impermeable for succinate (but see Zhunussova 2015 Am J Cancer Res for an exception). Incubation of mt-preparations by succinate alone may lead to accumulation of oxaloacetate, which is a potent inhibitor of Complex II (compare Succinate and rotenone). High activities of mtMalic enzyme (mtME) prevent accumulation of oxaloacetate.
Abbreviation: S
Reference: Gnaiger 2014 MitoPathways, Tretter 2016 Biochim Biophys Acta
MitoPedia topics:
Substrate and metabolite
Application in HRFR
S: Succinate (Succinate disodium salt, hexahydrate, C4H404Na2 * (H2O)6); Sigma S 2378, 100 g, store at RT; FW = 270.1
- Preparation of 1 M stock solution (dissolved in H2O)
- Weight 1.3505 g of succinate and dissolve in 3 mL H2O;
- Check pH and adjust to 7.0 if necessary with 1 M HCl (usually the pH is 7 without any adjustment);
- Transfer to 5 mL volumetric glass flask and adjust the final volume to 5 mL;
- Divide into 0.5 mL portions;
- Store frozen at -20 °C.
- O2k manual titrations MiPNet09.12 O2k-Titrations
- In the absence of CI-linked substrates, add the CI-inhibitor rotenone before addition of succinate, to avoid accumulation of oxaloacetate with subsequent inhibition of succinate dehydrogenase. See: Succinate and rotenone.
- When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm the stock solution in your hands if necessary.
- Titration volume: 20 µL using a 50 µL syringe (2 mL O2k-chamber).
- Final concentration: 10 mM.