Adaptation of an enzymatic fluorescence assay for L-glutamic acid decarboxylase.
The activity of L-glutamic acid decarboxylase (GAD) is commonly estimated by several radiometric methods, whereas a fluorimetric assay based on an enzymatic formation of NADPH as described by Y. Okada and C. Shimada [(1975) Brain Res. 98, 202-206] has been given little attention in biochemical and pharmacological investigations. A simple modification of this assay is presented to permit rapid and sensitive GAD measurements in unpurified tissue homogenates. This method, employing a linear NADPH standard curve, is demonstrated to be a valid assay system for a pharmacological approach using 3-mercaptopropionic acid.
Wolf
R
R
Max-Planck-Institute for Psychiatry, Munich, Federal Republic of Germany.
Klemisch
H
H
D015097:3-Mercaptopropionic Acid; D000818:Animals; D046508:Culture Techniques; D005453:Fluorescence; D005470:Fluorometry; D005968:Glutamate Decarboxylase / Q000032:analysis*; D007700:Kinetics; D009249:NADP / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000201:enzymology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D009249:NADP; D015097:3-Mercaptopropionic Acid; D005968:Glutamate Decarboxylase