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Difference between revisions of "Malonate"

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{{MitoPedia
{{MitoPedia
|abbr=Mna
|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]]
|info=[[MiPNet09.12]]
|type=Inhibitor ETS
}}
}}
{{Labeling
{{MitoPedia topics
|discipline=Mitochondrial Physiology
|mitopedia topic=Inhibitor
|enzymes=Complex II; Succinate Dehydrogenase
|kinetics=Inhibitor; Uncoupler
|instruments=Chemicals; Media
|type=Inhibitor ETS
}}
}}
 
__TOC__
== Application in [[HRR]] ==
== Application in [[HRR]] ==


'''Mna: Malonate''' (Malonic acid, CH<sub>2</sub>(COOH)<sub>2</sub>); Sigma M 1296, 5 g, store at -20 °C: FW = 800
=== 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"> '''Caution:''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>
<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): Recommended to prepare fresh every day
::::# 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 H<sub>2</sub>O to a final volume of 100 µL.


::1) weigh 20.8 mg of Malonic acid directly into a Eppendorf tube of appropriate volume
::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
::2) dissolve in 21 µl H20 + 79 µl 5 N KOH
::3) check pH - should be around pH of 6.0. 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 KOH (1 µl).


'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
::::* Titration volume: 5 µL using a 25 µL syringe (2 mL O2k-chamber).
::::* Final concentration: 5 mM.


::* Titration volume: 5 µl using a 10 µl syringe
===  Malonic acid and malonyl-CoA synthase ===
::* Final conc. in 2 ml O2k-chamber: 5 mM
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. - [[Witkowski 2011 J Biol Chem |»Bioblast link«]]

Revision as of 12:07, 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):
  1. Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
  2. Dissolve in 85 µL 5 M KOH and check pH.
  3. 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).
  4. 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«