title: Purification of L-glutamate decarboxylase from monkey brain.
abstract: Glutamate decarboxylase (GAD) is an enzyme that synthesizes gamma-aminobutyrate (GABA), a major inhibitory neurotransmitter in the central nervous system. Post-translational modification of GAD, such as N-terminal blockage, phosphorylation-dephosphorylation, and palmitoylation, is an important factor in the biological activity of GAD. In order to address the significance of post-translational events on GAD, we thought it crucial to obtain a non-recombinant form of GAD. In this study, we attempted to isolate GAD protein from the monkey brain, a model animal close to the human that has not been studied. Monkey brain was homogenized, fractionated with ammonium sulphate, and applied to a series of chromatographic steps, including hydrophobic, ion-exchange, and gel filtration. Purified GAD showed a single band on SDS-PAGE, and the enzyme was found to have a molecular weight of 61,000 and exhibited 1,100 nmol/min/mg of specific activity. It had an optimal pH of 7 and optimal thermal stability at 40 degrees C.
authors.affiliation: Laboratory of Applied Microbiology and Biochemistry, Nara Women's University, Nara, Japan.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Inoue
authors.forename: Kenji
authors.initials: K
authors.lastname: Ishii
authors.forename: Mizuo
authors.initials: M
authors.lastname: Miyazaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ueno
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D005968:Glutamate Decarboxylase / Q000032:analysis / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008251:Macaca; D008297:Male; D013696:Temperature
chemical_list: D005968:Glutamate Decarboxylase