Assay and properties of glutamic acid decarboxylase in homogenates of crayfish nervous tissue

Abstract

The activity of glutamic acid decarboxylase (GAD) was measured in homogenates of crayfish nervous tissue. Radioactive GABA and CO2 were formed from radioactive glutamic acid in approximately equimolar amounts. Product formation was linear for 9.5 hr at 11-32 degrees C with about 1-30 micrograms homogenate protein. Enzyme activity remained high at pH 7-10 but declined steeply above pH 10.5 and below pH 7. Enzyme activity was stimulated by pyridoxal phosphate, 2-mercaptoethanol, and potassium phosphate; at higher than optimal concentrations of each the activity was reduced. Sodium phosphate altered the stimulatory effect of potassium phosphate. Crayfish GAD behaves like a typical neural GAD but is distinguishable biochemically from GAD of other species.