Difference between revisions of "Malonate"

Revision as of 13:07, 17 March 2019

high-resolution terminology - matching measurements at high-resolution

Malonate

Description

Malonate (malonic acid) is a competitive inhibitor of succinate dehydrogenase (Complex II). Malonate is a substrate of malonyl-CoA synthase.

Abbreviation: Mna

Reference: MiPNet09.12

MitoPedia topics: Inhibitor

Application in HRR

Preparation and titration

Mna: Malonic acid ( CH₂(COOH)₂); Sigma M 1296, 5 g, store at R.T.; FW = 104.06.

Recommendation: Prepare fresh everyday!

Preparation of 2 M stock solution (dissolved in H20):

Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
Dissolve in 85 µL 5 M KOH and check pH.
Titrate small volumes (2 µL) of 5 M KOH until you reach a pH of 6.0 (pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very small excess KOH).
Add H₂O to a final volume of 100 µL.

O2k manual titrations MiPNet09.12 O2k-Titrations

Titration volume: 5 µL using a 25 µL syringe (2 mL O2k-chamber).
Final concentration: 5 mM.

Malonic acid and malonyl-CoA synthase

Communicated by Garcia-Souza LF 2016-11-11

Activity of mt-malonyl-CoA synthase (Witkowski et al 2011) in a mt-preparation may cause stimulation of respiration after titration of malonate (unpublished observation).

Reference

Witkowski A, Thweatt J, Smith S (2011) Mammalian ACSF3 protein is a malonyl-CoA synthetase that supplies the chain extender units for mitochondrial fatty acid synthesis. J Biol Chem 286:33729-36. - »Bioblast link«

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=Mna
-|description='''Malonate''' (malonic acid) is a competitive inhibitor of [[Succinate dehydrogenase]] ([[Complex II]]).
+|description='''Malonate''' (malonic acid) is a competitive inhibitor of [[succinate dehydrogenase]] ([[Complex II]]). Malonate is a substrate of [[malonyl-CoA synthase]].
 |info=[[MiPNet09.12]]
-|type=Inhibitor ETS
 }}
 {{MitoPedia topics
 |mitopedia topic=Inhibitor
-|type=Inhibitor ETS
 }}
+__TOC__
 == Application in [[HRR]] ==
-'''Mna: Malonate''' (Malonic acid, CH<sub>2</sub>(COOH)<sub>2</sub>); Sigma M 1296, 5 g, store at R.T.; FW = 104.06
+=== Preparation and titration ===
+::: '''Mna: Malonic acid''' ( CH<sub>2</sub>(COOH)<sub>2</sub>); Sigma M 1296, 5 g, store at R.T.; FW = 104.06.
 <span style="color:#8B008B"> '''Recommendation:''' Prepare fresh everyday!</span>
+::: '''Preparation of 2 M stock solution''' (dissolved in H20):
-'''Preparation of 2 M stock solution''' (dissolved in H20):
+::::# Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
+::::# Dissolve in 85 µL 5 M KOH and check pH.
-::1) weigh 20.8 mg of Malonic acid directly into a Eppendorf tube of appropriate volume (final volume = 100 µl)
+::::# Titrate small volumes (2 µL) of 5 M KOH until you reach a pH of 6.0 (pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very small excess KOH).
-::2) dissolve in 20 µl H<sub>2</sub>O
+::::# Add H<sub>2</sub>O to a final volume of 100 µL.
-::3) add 70 µl 5 N KOH and check pH
-::4) titrate small amounts (2 µl) of 5 N KOH until you reach a pH of 6.0 (approx. 4 titration steps). A pH of 7.0 would be ideal, however the pH jumps from pH 6.0 to pH 10.0 with only very little amounts of excess KOH.
-::5) add H<sub>2</sub>O to a final volume of 100 µl
+::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
+::::* Titration volume: 5 µL using a 25 µL syringe (2 mL O2k-chamber).
+::::* Final concentration: 5 mM.
-'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
+===  Malonic acid and malonyl-CoA synthase ===
+ Communicated by [[Garcia-Souza LF]] 2016-11-11
+:::: Activity of mt-[[malonyl-CoA synthase]] (Witkowski et al 2011) in a mt-preparation may cause stimulation of respiration after titration of malonate (unpublished observation).
-::* Titration volume: 5 µl using a 25 µl syringe
+:::: Reference
-::* Final conc. in 2 ml O2k-chamber: 5 mM
+::::* Witkowski A, Thweatt J, Smith S (2011) Mammalian ACSF3 protein is a  malonyl-CoA synthetase that supplies the chain extender units for  mitochondrial fatty acid synthesis. J Biol Chem 286:33729-36. - [[Witkowski 2011 J Biol Chem |»Bioblast link«]]