Determination of glutamate decarboxylase by high-performance liquid chromatography

被引：46

作者：

Rossetti, V

Lombard, A

机构：

来源：

JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1996年 / 681卷 / 01期

关键词：

glutamate decarboxylase; enzymes;

D O I：

10.1016/0378-4347(96)88202-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An improved method for the determination of glutamate decarboxylase (GAD) activity is described. The enzyme was evaluated by incubation with glutamic acid (L-Glu) in the presence of pyridoxal 5'-phosphate (PLP): the gamma-aminobutyric acid (GABA) formed was derivatized to PTC-GABA; the latter was subsequently separated and assayed by isocratic HPLC (LiChrospher RP-18 column; isocratic elution with pH 5.8 acetate buffer in acetonitrile-water) with UV absorbance detection at 254 nm. The method described is a sensitive, reproducible and specific assay useful for following variations of GAD activity in vitro; this assay was subsequently used for the evaluation of GAD activity variations after irradiation with low doses of He-Ne laser radiation.

引用

页码：63 / 67

页数：5

共 14 条

[1] REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR DETERMINATION OF BRAIN GLUTAMATE-DECARBOXYLASE SUITABLE FOR USE IN KINETIC-STUDIES [J].