Malonate: Difference between revisions
From Bioblast
No edit summary |
(Added instructions for the sV-chamber) |
||
| Line 11: | Line 11: | ||
::::<span style="color:#8B008B"> '''Recommendation:''' Prepare fresh everyday!</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) 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 20: | Line 21: | ||
:::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | :::» '''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. | ::::* Final concentration: 5 mM. | ||
=== Malonic acid and malonyl-CoA synthase === | === Malonic acid and malonyl-CoA synthase === | ||
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«
- Reference
MitoPedia topics: Inhibitor
