A rapid colorimetric assay for gamma-glutamyl hydrolase (conjugase).
A simple procedure for the measurement of gamma-glutamyl hydrolase (conjugase) activity is described. Glutamic acid released from pteroylpenta-gamma-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-gamma-glutamate substrate by conjugase, conversion of glutamate to alpha-ketoglutarate by L-glutamate dehydrogenase and colorimetric measurement of the BDBH derivative of alpha-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.
Department of Biochemical Pharmacology, State University of New York, Buffalo 14260.
P V
PV
Wagh
T I
TI
Kalman
D000818:Animals; D001577:Benzophenones; D002417:Cattle; D002645:Chickens; D003124:Colorimetry / Q000379:methods; D005492:Folic Acid / Q000378:metabolism; D005971:Glutamates / Q000032:analysis; D018698:Glutamic Acid; D006868:Hydrolysis; D007668:Kidney / Q000201:enzymology; D010179:Pancreas / Q000201:enzymology; D011624:Pteroylpolyglutamic Acids / Q000378:metabolism; D013552:Swine; D011623:gamma-Glutamyl Hydrolase / Q000032:analysis* / Q000378:metabolism
D001577:Benzophenones; D005971:Glutamates; D011624:Pteroylpolyglutamic Acids; C036907:pteroylpentaglutamic acid; D018698:Glutamic Acid; C014682:Michler's ketone; D005492:Folic Acid; D011623:gamma-Glutamyl Hydrolase