Succinate: Difference between revisions

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Succinate

Description

Succinic acid, C₄H₆O₄, (butanedioic acid) is a dicarboxylic acid which occurs under physiological conditions as the anion succinate^2-, S, with pK_a1 = 4.2 and pK_a2 = 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

Succinate, S. From Gnaiger 2014 MitoPathways.

Application in HRR

S: Succinate (Sodium succinate dibasic hexahydrate; C₄H₄0₄Na₂ * 6H₂O), Sigma-Aldrich: S2378, store at RT, CAS: 6106-21-4, M = 270.1 g·mol^-1

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.

Preparation of 1 M stock solution:

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 (about 65 µL will be needed);
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

Titration volume (2-mL O2k-chamber): 20 µL using a 50 µL Hamilton syringe.
Titration volume (0.5-mL O2k-chamber): 5 µL using a 25 µL Hamilton syringe
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.

MitoPedia topics: Substrate and metabolite

@@ Line 9: / Line 9: @@
 [[File:S.jpg|400px|thumb|Succinate, S. From [[Gnaiger 2014 MitoPathways]].]]
 == Application in [[HRR]] ==
+{{Chemical_description
+|abbr=S
+|trivial name=Succinate
+|complete name=Sodium succinate dibasic hexahydrate
+|chem formula= C<sub>4</sub>H<sub>4</sub>0<sub>4</sub>Na<sub>2</sub> * 6H<sub>2</sub>O
-:::'''S: Succinate''' (Succinate disodium salt, hexahydrate, C<sub>4</sub>H<sub>4</sub>0<sub>4</sub>Na<sub>2</sub> * (H<sub>2</sub>O)<sub>6</sub>; Sigma S 2378, 100 g, store at RT; M = 270.1 g·mol<sup>-1</sup>)
+|molar mass=270.1
+|vendor=Sigma-Aldrich
+|product number=S2378
+|store at=RT
+|sensitivity=
+|cas=6106-21-4
+|h statements=
+|h info=
+}}<!--:::'''S: Succinate''' (Succinate disodium salt, hexahydrate, C<sub>4</sub>H<sub>4</sub>0<sub>4</sub>Na<sub>2</sub> * (H<sub>2</sub>O)<sub>6</sub>; Sigma S 2378, 100 g, store at RT; M = 270.1 g·mol<sup>-1</sup>)-->
 :::: 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.