Difference between revisions of "Glutamate"
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'''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. | '''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] | |info=[http://www.oroboros.at/?Gnaiger_2012_MitoPathways Gnaiger 2012 MitoPathways] | ||
}} | }} | ||
{{MitoPedia methods|type=Respiration | {{MitoPedia methods|type=Respiration | ||
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{{MitoPedia topics | {{MitoPedia topics | ||
|mitopedia topic=Substrate and metabolite | |mitopedia topic=Substrate and metabolite | ||
}} | }} | ||
== Application in [[HRR]] == | == Application in [[HRR]] == | ||
'''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) | ||
:: | ::::# Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 ml volumetric glass flask. | ||
:: | ::::# Add 3 ml H<sub>2</sub>O. | ||
:: | ::::# Adjust pH with 5 N KOH to pH 7.0. | ||
:: | ::::# Adjust final volume to 5 ml and divide into 0.5 ml portions. | ||
:: | ::::# Store at -20 °C. | ||
'''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | :::: '''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 09:35, 9 April 2016
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)
- Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 ml volumetric glass flask.
- Add 3 ml H2O.
- Adjust pH with 5 N KOH to pH 7.0.
- Adjust final volume to 5 ml and divide into 0.5 ml portions.
- Store 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.