Assay for aliphatic amino acid decarboxylases by high-performance liquid chromatography

Kochhar, Sunil; Mehta, Perdeep K.; Christen, Philipp

doi:10.1016/0003-2697(89)90221-2

Cited by 11 publications

(1 citation statement)

References 6 publications

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“…The filtrate was diluted with one volume of HPLC medium and loaded at room temperature onto a Nucleosil 100-5C18 reversed phase column (Macherey-Nagel, 4.8 × 250 mm) attached to an HPLC system (Beckman). The column was eluted isocratically at a flow rate of 1 ml/min, with 13 mM trifluoroacetic acid containing 1 mM 1-octanesulfonate as a mobile phase (43). Elution of glutamate and other compounds was detected spectrophotometrically at 214 nm.…”

Section: Methodsmentioning

confidence: 99%

Synaptic Vesicle-bound Pyruvate Kinase can Support Vesicular Glutamate Uptake

2008

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Glucose metabolism is essential for normal brain function and plays a vital role in synaptic transmission. Recent evidence suggests that ATP synthesized locally by glycolysis, particularly via glyceraldehyde 3-phosphate dehydrogenase/3-phosphoglycerate kinase, is critical for synaptic transmission. We present evidence that ATP generated by synaptic vesicle-associated pyruvate kinase is harnessed to transport glutamate into synaptic vesicles. Isolated synaptic vesicles incorporated [ 3 H]glutamate in the presence of phosphoenolpyruvate (PEP) and ADP. Pyruvate kinase activators and inhibitors stimulated and reduced PEP/ADP-dependent glutamate uptake, respectively. Membrane potential was also formed in the presence of pyruvate kinase activators. "ATP-trapping" experiments using hexokinase and glucose suggest that ATP produced by vesicleassociated pyruvate kinase is more readily used than exogenously added ATP. Other neurotransmitters such as GABA, dopamine, and serotonin were also taken up into crude synaptic vesicles in a PEP/ADP-dependent manner. The possibility that ATP locally generated by glycolysis supports vesicular accumulation of neurotransmitters is discussed.

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Section: Methodsmentioning

confidence: 99%