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Difference between revisions of "Succinate"

From Bioblast
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::::* 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 µL using a 50 µL syringe (2 mL O2k-chamber).
::::* Titration volume: 20 µL using a 50 µL syringe (2 mL O2k-chamber).
::::* Final concentration: 10 mM.
::::* Final concentration: 10 mM. Note: The concentration of succinate may be increased up to 50 mM after [[rotenone]] to compensate for the inhibitory effect of [[malate]].     


== [[SUITbrowser]] question: Succinate pathway ==
== [[SUITbrowser]] question: Succinate pathway ==
:::: The [https://suitbrowser.oroboros.at/ SUITbrowser] can be used to find the best SUIT protocols to analyze the succinate pathway, among other research questions.
:::: The [https://suitbrowser.oroboros.at/ SUITbrowser] can be used to find the best SUIT protocols to analyze the succinate pathway, among other research questions.

Revision as of 10:48, 8 November 2019


high-resolution terminology - matching measurements at high-resolution


Succinate

Description

Succinic acid

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 mt-Malic enzyme (mtME) prevent accumulation of oxaloacetate.

Abbreviation: S

Reference: Gnaiger 2019 MitoPathways, Tretter 2016 Biochim Biophys Acta


MitoPedia topics: Substrate and metabolite 



Succinate, S. From Gnaiger 2014 MitoPathways.

Application in HRR

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:
  1. Weight 1.3505 g of succinate and dissolve in 3 mL H2O;
  2. Check pH and adjust to 7.0 if necessary with 1 M HCl (usually the pH is 7 without any adjustment);
  3. Transfer to 5 mL volumetric glass flask and adjust the final volume to 5 mL;
  4. Divide into 0.5 mL portions;
  5. 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. Note: The concentration of succinate may be increased up to 50 mM after rotenone to compensate for the inhibitory effect of malate.

SUITbrowser question: Succinate pathway

The SUITbrowser can be used to find the best SUIT protocols to analyze the succinate pathway, among other research questions.