Difference between revisions of "Succinate"
From Bioblast
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|abbr=S | |abbr=S | ||
|description=[[File:Succinic_acid.jpg|left|100px|Succinic acid]] | |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>, S''', 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>, 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 mitochondrial membrane by the [[dicarboxylate carrier]]. 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. | ||
Succinate is formed in the [[TCA cycle]], and is a substrate of [[Complex II | | |info=[[Gnaiger 2014 MitoPathways]] | ||
|info=[[Gnaiger | |||
|type=Respiration | |type=Respiration | ||
}} | }} | ||
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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): | ||
::1 | ::1. Weigh 1.3505 g of succinate and dissolve in 3 ml H<sub>2</sub>O. | ||
::2 | ::2. Check pH and adjust to 7.0 if necessary with 1 N HCl (usually the pH is 7 without any adjustment). | ||
::3 | ::3. Transfer to 5 ml volumetric glass flask and adjust the final volume to 5 ml. | ||
::4 | ::4. Divide into 0.5 ml portions. | ||
::5 | ::5. Store frozen at -20 °C. | ||
'''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | '''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::* In the absence 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 succiante 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 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. | ||
Revision as of 01:08, 10 March 2015
- high-resolution terminology - matching measurements at high-resolution
Succinate
Description
Succinic acid, C4H6O4, 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 mitochondrial membrane by the dicarboxylate carrier. 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
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 (dissolved in H2O):
- 1. Weigh 1.3505 g of succinate and dissolve in 3 ml H2O.
- 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
- 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. 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.
