Difference between revisions of "Succinate"

Latest revision as of 15:06, 17 December 2022

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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 in incubations with succinate without rotenone.

Abbreviation: S

Reference: Gnaiger 2020 BEC MitoPathways, Tretter 2016 Biochim Biophys Acta

Succinate, S. From Gnaiger 2020 BEC 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 2: / Line 2: @@
 |abbr=S
 |description=[[File:Succinic_acid.jpg|left|100px|Succinic acid]]
-'''Succinic acid''', C<sub>4</sub>H<sub>6</sub>O<sub>4</sub>, occurs under physiological conditions as the anion '''succinate<sup>2-</sup>, M''', with ''p''K<sub>a1</sub> = 4.2 and ''p''K<sub>a2</sub> = 5.6.
+'''Succinic acid''', C<sub>4</sub>H<sub>6</sub>O<sub>4</sub>, (butanedioic acid) is a dicarboxylic acid which occurs under physiological conditions as the anion '''succinate<sup>2-</sup>, S''', with ''p''K<sub>a1</sub> = 4.2 and ''p''K<sub>a2</sub> = 5.6. Succinate is formed in the [[TCA cycle]], and is a substrate of [[Complex II |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 in incubations with succinate without rotenone.
-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 mitochondrial membrane by the [[dicarboxylate carrier]].
+|info=[[Gnaiger 2020 BEC MitoPathways]], [[Tretter 2016 Biochim Biophys Acta]]
-|info=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways]
-|type=Respiration
 }}
-{{MitoPedia methods|type=Respiration
+__TOC__
-}}
-{{MitoPedia topics
+[[File:S.jpg|400px|thumb|Succinate, S. From [[Gnaiger 2020 BEC MitoPathways]].]]
-|mitopedia topic=Substrate and metabolite
+== Application in [[HRR]] ==
-|type=Respiration
+{{Chemical_description
-}}
+|abbr=S
-== Application in [[HRR]]  ==
+|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
+|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.
+:::: '''Preparation of 1 M stock solution:'''
+::::# 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 (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.
-'''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; FW = 270.1
+:::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
-'''Preparation of 1 M stock solution''' (dissolved in H<sub>2</sub>O):
+::::* 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]].
-::1) Weigh 1.3505 g of succinate and dissolve in 3 ml H<sub>2</sub>O.
-::2) Check pH and adjust to 7.0 if necessary with 1 N 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.
-'''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
+== [[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.
-::* 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 succiante dehydrogenase.
+{{MitoPedia topics
-::* When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm in your hands if necessary.
+|mitopedia topic=Substrate and metabolite
-::* Titration volume: 20 µl using a 50 µl syringe (2 ml O2k-chamber).
+}}
-::* Final concentration: 10 mM.