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Glutamate

From Bioblast


high-resolution terminology - matching measurements at high-resolution


Glutamate

Description

Glutamate as the sole substrate is transported by the electroneutral glutamate-/OH- exchanger, and is oxidized via glutamate dehydrogenase in the mitochondrial matrix. Ammonia can pass freely through the mitochondrial membrane. See also Glutamate-aspartate carrier.

Abbreviation: G

Reference: MiPNet11.04; MiPNet09.12


Labels:



Enzyme: Inner mtMembrane Transporter"Inner mtMembrane Transporter" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property.  Regulation: Substrate; Glucose; TCA Cycle"Substrate; Glucose; TCA Cycle" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property. 


HRR: Chemicals; Media"Chemicals; Media" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 



Application in HRR

G: Glutamate (L-Glutamic acid, sodium salt, C5H8NO4Na; contains 1 mol/mol H2O); Sigma G 1626, 100 g, store at R.T.: FW = 187.1 or 169.1 anhydrous)

Preparation of 2 M stock solution (dissolved in H2O):

1) weigh 1.871 g of L-Glutamic acid and transfer to 5 mL volumetric glass flask
2) add 3 ml H2O (pH = 6)
3) neutralize with 5 N KOH (approx. 50 ยตl)
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 ยฐC

Oxygraph-2k Manual Titrations: MiPNet09.12

  • Titration volume: 10 ยตl using a 25 ยตl syringe
  • Final conc. in 2 ml O2k-chamber: 10 mM