Difference between revisions of "Succinate"

Revision as of 20:41, 1 June 2013

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Succinate

Description

Succinate^2- 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.

Abbreviation: S

Reference: Gnaiger 2012 MitoPathways

MitoPedia topics: Substrate and metabolite

Application in HRR

S: Succinate (Succinate disodium salt, hexahydrate, C₄H₄0₄Na₂ * (H₂O)₆); Sigma S 2378, 100 g, store at R.T.; 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 0.2 ml portions.

5) Store frozen at -20 °C.

Oxygraph-2k Manual Titrations: MiPNet09.12

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.
When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm in your hands if necessary.
Titration volume: 20 µl using a 50 µl syringe (2 ml O2k-chamber).
Final concentration: 10 mM.

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=S
-|description='''Succinate<sup>2-</sup>''' 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.
+|description='''Succinate<sup>2-</sup>''' 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=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways], [[MiPNet12.11]]
+|info=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways]
 |type=Respiration
 }}
@@ Line 17: / Line 17: @@
 '''Preparation of 1 M stock solution''' (dissolved in H<sub>2</sub>O):
-::1) weigh 1.3505 g of Succinate and dissolve in 3 ml H<sub>2</sub>O
+::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)
+::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
+::3) Transfer to 5 ml volumetric glass flask and adjust the final volume to 5 ml.
-::4) divide into 0.2 ml portions
+::4) Divide into 0.2 ml portions.
-::5) store frozen at -20 °C
+::5) Store frozen at -20 °C.
 '''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]]
-::* In the absence of CI-linked substrates, add rotenone before addition of succinate, to avoid accumulation of oxaloacetate with subsequent Inhibition of succiante Dehydrogenase.
+::* 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.
-::* Titration volume: 20 µl using a 50 µl syringe (2 ml O2k-chamber)
+::* When keeping the succinate stock solution on ice, check for complete solubilization of succinate and warm in your hands if necessary.
-::* Final concentration: 10 mM
+::* Titration volume: 20 µl using a 50 µl syringe (2 ml O2k-chamber).
+::* Final concentration: 10 mM.