Difference between revisions of "Glutamate"
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''' | '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::* Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber). | ::* Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber). | ||
::* Final concentration: 10 mM. | ::* Final concentration: 10 mM. |
Revision as of 14:17, 16 June 2015
Description
Glutamic acid, C5H9NO4, is an amino acid which occurs under physiological conditions mainly as the anion glutamate-, G, with pKa1 = 2.1, pKa2 = 4.07 and pKa3 = 9.47. Glutamate+malate is a CI-linked substrate combination, when glutamate is transported into the mt-matrix via the glutamate-aspartate carrier and reacts with oxaloacetate in the transaminase reaction to form aspartate and oxoglutarate. Glutamate as the sole substrate is transported by the electroneutral glutamate-/OH- exchanger, and is oxidized in the mitochondrial matrix by glutamate dehydrogenase to α-ketoglutarate ( 2-oxoglutarate). Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane.
Abbreviation: G
Reference: Gnaiger 2012 MitoPathways
MitoPedia topics: Substrate and metabolite
Application in HRR
G: Glutamate (L-Glutamic acid, monosodium salt hydrate, C5H8NO4Na; contains 1 mol H2O /mol); Sigma G 1626, 100 g, store at R.T.; FW = 169.1)
Preparation of 2 M stock solution (dissolved in H2O)
- 1) Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to 5 mL volumetric glass flask.
- 2) Add 3 ml H2O.
- 3) Check pH and adjust with 5N KOH to 7.
- 4) Adjust final volume to 5 ml and divide into 0.5 ml portions.
- 5) Store frozen at -20 °C.
O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber).
- Final concentration: 10 mM.