Difference between revisions of "Malonate"

Revision as of 09:29, 23 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:

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 (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:

Weigh 10.4 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 200 µL).
Dissolve in 170 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 200 µL.

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synthase (Witkowski et al 2011) in a mt-preparation may cause stimulation of respiration after titration of malonate (unpublished observation).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

@@ Line 11: / Line 11: @@
 ::::<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) for '''2-mL O2k-chamber''':
 ::::# Weigh 20.8 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 100 µL).
@@ Line 20: / Line 21: @@
 :::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
-::::* Titration volume: 5 µL using a 25 µL syringe (2 mL O2k-chamber).
+::::* 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''':
+::::# Weigh 10.4 mg malonic acid into a Eppendorf tube of appropriate volume (for final volume of 200 µL).
+::::# Dissolve in 170 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 200 µL.
+:::» '''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 ===