Difference between revisions of "Glutamate"
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{{MitoPedia | {{MitoPedia | ||
|abbr=G | |abbr=G | ||
|description=''' | |description=[[File:Glutamic_acid.jpg|left|80px|Glutamic acid]] | ||
'''Glutamic acid''', C<sub>5</sub>H<sub>9</sub>NO<sub>4</sub>, is an amino acid which occurs under physiological conditions mainly as the anion '''glutamate<sup>-</sup>, G''', with ''p''K<sub>a1</sub> = 2.1, ''p''K<sub>a2</sub> = 4.07 and ''p''K<sub>a3</sub> = 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<sup>-</sup>/OH<sup>-</sup> exchanger, and is oxidized in the mitochondrial matrix by [[glutamate dehydrogenase]] to α-ketoglutarate ([[oxoglutarate | 2-oxoglutarate]]). Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane. | |||
|info=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways], [[MiPNet09.12]] | |info=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways], [[MiPNet09.12]] | ||
|type=Respiration | |type=Respiration | ||
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'''G: Glutamate''' (L-Glutamic acid, monosodium salt hydrate, C<sub>5</sub>H<sub>8</sub>NO<sub>4</sub>Na; contains 1 mol H<sub>2</sub>O /mol); Sigma G 1626, 100 g, store at R.T.; FW = 169.1) | '''G: Glutamate''' (L-Glutamic acid, monosodium salt hydrate, C<sub>5</sub>H<sub>8</sub>NO<sub>4</sub>Na; contains 1 mol H<sub>2</sub>O /mol); Sigma G 1626, 100 g, store at R.T.; FW = 169.1) | ||
'''Preparation of 2 M stock solution''' (dissolved in H<sub>2</sub>O) | '''Preparation of 2 M stock solution''' (dissolved in H<sub>2</sub>O) | ||
::1) | ::1) Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to 5 mL volumetric glass flask. | ||
::2) | ::2) Add 3 ml H<sub>2</sub>O (pH = 6). | ||
::3) | ::3) Neutralize with 5 N KOH. | ||
::4) | ::4) Check pH and adjust to 7 if necessary. | ||
::5) | ::5) Adjust final volume to 5 ml and divide into 0.5 ml portions. | ||
::6) | ::6) Store frozen at -20 °C. | ||
'''Oxygraph-2k | '''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::* Titration volume: 10 µl using a 25 µl syringe | ::* Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber). | ||
::* Final | ::* Final concentration: 10 mM. | ||
Revision as of 10:22, 15 April 2014
- high-resolution terminology - matching measurements at high-resolution
Glutamate
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, MiPNet09.12
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 (pH = 6).
- 3) Neutralize with 5 N KOH.
- 4) Check pH and adjust to 7 if necessary.
- 5) Adjust final volume to 5 ml and divide into 0.5 ml portions.
- 6) Store frozen at -20 °C.
Oxygraph-2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 10 µl using a 25 µl syringe (2 ml O2k-chamber).
- Final concentration: 10 mM.
