Abstract
A simple procedure for the measurement of γ-glutamyl hydrolase (conjugase) activity is described. Glutamic acid released from pteroylpenta-γ-glutamate by hog kidney and chicken pancreas conjugases was quantitated using the dye 4,4′-bis(dimethylamino)benzophenone hydrazone. The procedure involves hydrolysis of the folylpoly-γ-glutamate substrate by conjugase, conversion of glutamate to α-ketoglutarate by l-glutamate dehydrogenase and colorimetric measurement of the BDBH derivative of α-ketoglutarate. The release of as little as one nmol of glutamic acid from the substrate can be measured by this procedure, which is well suited for the assay of a variety of conjugase preparations. In addition, the method should provide a general assay for the enzymatic hydrolysis of various folate and antifolate polyglutamates.
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