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Difference between revisions of "Malonate"

From Bioblast
(Added instructions for the sV-chamber)
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:::: '''Preparation of 2 M stock solution''' (dissolved in H20) for '''2-mL O2k-chamber''':  
:::: '''Preparation of 2 M stock solution''' (dissolved in H<sub>2</sub>0) for '''2-mL O2k-chamber''':  


::::# Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
::::# Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
Line 25: Line 25:




:::: '''Preparation of 0.5 M stock solution''' (dissolved in H20) for '''0.5-mL O2k-chamber''':  
:::: '''Preparation of 0.5 M stock solution''' (dissolved in H<sub>2</sub>0) for '''0.5-mL O2k-chamber''':  


::::# Weigh 10.4 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 200 µL).
::::# Take 50 µL of 2 M stock solution
::::# Dissolve in 170 µL 5 M KOH and check pH.
::::# Add 150 µL H<sub>2</sub>O
::::# 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 200 µL.


:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]

Revision as of 13:22, 27 October 2020


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


Application in HRR

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


Preparation of 0.5 M stock solution (dissolved in H20) for 0.5-mL O2k-chamber:
  1. Take 50 µL of 2 M stock solution
  2. Add 150 µL H2O
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (0.5-mL O2k-chamber): 5 µL using a 25 µL syringe.
  • Final concentration: 5 mM.


Malonic acid and malonyl-CoA synthase

Communicated by Garcia-Souza Luiz F 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«

MitoPedia topics: Inhibitor