The measurement of gamma-glutamyl hydrolase (conjugase) activity in rat brain

descriptionPublicationkeyboard_double_arrow_right Article 01 Feb 1988 English Publisher:Springer Science and Business Media LLCJournal:Neurochemical Research, volume 13, pages 147-151 (issn: 0364-3190, eissn: 1573-6903,

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Authors: D M, Hartley; S R, Snodgrass; P A, Bradshaw;

doi: 10.1007/bf00973326

pmid: 3362292

The measurement of gamma-glutamyl hydrolase (conjugase) activity in rat brain

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Abstract

An assay using the artificial substrate, 2,4-diamino-10-methyl-pteroylglutamyl-gamma-glutamate (MTX-G1), was developed to measure gamma-glutamyl hydrolase (conjugase), which hydrolyzes folylpolyglutamates. This assay allows us to: 1) measure conjugase for the first time in rat brain and 2) measure conjugase in a reliable, sensitive and inexpensive manner. The MTX-binding assay results were compared to samples analyzed by HPLC and found to vary by only 13%. The artificial substrate, MTX-G1, had a lower rate of hydrolysis than pteroylglutamyl-gamma-glutamate (Pte-G2), 70.7 +/- 0.64 and 92.6 +/- 0.22 nmoles/hr/mg protein respectively. Conjugase was semi-purified 24 fold in H2O and found to have a pH optimum of 5.0.

Related Organizations

University of California System
United States
University of Southern California
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Children's Hospital of Los Angeles
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Stanford University
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Keywords

Male, Cysteine Endopeptidases, Animals, Brain, gamma-Glutamyl Hydrolase, Hydrogen-Ion Concentration, Chromatography, High Pressure Liquid, Rats

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