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Difference between revisions of "Glutamate"

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{{MitoPedia
{{MitoPedia
|abbr=G
|abbr=G
|description='''Glutamate''' as the sole substrate is transported by the electroneutral glutamate-/OH- exchanger, and is oxidized in the mitochondrial matrix via [http://en.wikipedia.org/wiki/Glutamate_dehydrogenase glutamate dehydrogenase] to α-Ketoglutarate. Ammonia (the byproduct of the reaction) can pass freely through the mitochondrial membrane. See also [[Glutamate-aspartate carrier]].
|description=[[File:Glutamic_acid.jpg|left|100px|Glutamic acid]]
|info=[[MiPNet11.04]]; [[MiPNet09.12]]
'''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 substrate combination supporting an N-linked pathway control state, 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]]), representing the [[glutamate-anaplerotic pathway control state]]. Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane.
|type=Respiration
|info=[[Gnaiger 2020 BEC MitoPathways]]
}}
{{MitoPedia topics
|mitopedia topic=Substrate and metabolite
|type=Respiration
}}
}}
[[File:G.jpg|right|240px|link=Glutamate-anaplerotic pathway control state|G]]
__TOC__
== Application in [[HRR]] ==
== Application in [[HRR]] ==
{{Chemical_description
|abbr=G
|trivial name=Glutamate
|complete name=L-Glutamic acid, monosodium salt hydrate
|chem formula=C&#8325;H&#8328;NNaO&#8324; · xH&#8322;O
|molar mass=169.1
|vendor=Sigma-Aldrich
|product number=G1626
|store at=RT
|sensitivity=
|cas=142-47-2
|h statements=
|h info=
}}<!--::: '''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.; M = 169.1 g·mol<sup>-1</sup>)-->


'''G: Glutamate''' (L-Glutamic acid, sodium salt, C<sub>5</sub>H<sub>8</sub>NO<sub>4</sub>Na; contains 1 mol/mol H<sub>2</sub>O); Sigma G 1626, 100 g, store at R.T.: FW = 187.1 or 169.1 anhydrous)
:::: '''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 4 mL H<sub>2</sub>O.
::::# Check pH and adjust to 7.0 if necessary with 5 M KOH (usually the pH is 7 without any adjustment);
::::# Adjust final volume to 5 mL and divide into 0.5 mL portions.
::::# Store at -20 °C.


::1) weigh 1.871 g (or 1.691 g of anhydrous) L-Glutamic acid and transfer to 5 mL volumetric glass flask
:::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
::2) add 3 ml H<sub>2</sub>O (pH = 6)
::3) neutralize with 5 N KOH (approx. 50 µl)
::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]]
::::* Titration volume ('''2-mL O2k-chamber'''): 10 µL using a 25 µL Hamilton syringe.
::::* Titration volume ('''0.5-mL O2k-chamber'''): 2.5 µL using a 10 µL Hamilton syringe
::::* Final concentration: 10 mM.


::* Titration volume: 10 µl using a 25 µl syringe
 
::* Final conc. in 2 ml O2k-chamber: 10 mM
{{MitoPedia topics
|mitopedia topic=Substrate and metabolite
}}

Revision as of 10:12, 9 June 2021


high-resolution terminology - matching measurements at high-resolution


Glutamate

Description

Glutamic acid

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 substrate combination supporting an N-linked pathway control state, 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), representing the glutamate-anaplerotic pathway control state. Ammonia (the byproduct of the reaction) passes freely through the mitochondrial membrane.

Abbreviation: G

Reference: Gnaiger 2020 BEC MitoPathways

G

Application in HRR

G: Glutamate (L-Glutamic acid, monosodium salt hydrate; C₅H₈NNaO₄ · xH₂O), Sigma-Aldrich: G1626, store at RT, CAS: 142-47-2, M = 169.1 g·mol-1


Preparation of 2 M stock solution (dissolved in H2O)
  1. Weigh 1.691 g L-Glutamic acid, monosodium salt hydrate, and transfer to a 5 mL volumetric glass flask.
  2. Add 4 mL H2O.
  3. Check pH and adjust to 7.0 if necessary with 5 M KOH (usually the pH is 7 without any adjustment);
  4. Adjust final volume to 5 mL and divide into 0.5 mL portions.
  5. Store at -20 °C.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 10 µL using a 25 µL Hamilton syringe.
  • Titration volume (0.5-mL O2k-chamber): 2.5 µL using a 10 µL Hamilton syringe
  • Final concentration: 10 mM.


MitoPedia topics: Substrate and metabolite