一种用于γ-谷氨酰水解酶（结合酶）的快速比色测定法。
A rapid colorimetric assay for gamma-glutamyl hydrolase (conjugase).
作者信息
Wagh P V, Kalman T I
机构信息
Department of Biochemical Pharmacology, State University of New York, Buffalo 14260.
出版信息
Anal Biochem. 1992 Nov 15;207(1):1-5. doi: 10.1016/0003-2697(92)90490-x.
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.
本文描述了一种测量γ-谷氨酰水解酶（结合酶）活性的简单方法。使用染料4,4'-双（二甲基氨基）二苯甲酮腙对猪肾和鸡胰结合酶从蝶酰五-γ-谷氨酸释放的谷氨酸进行定量。该方法包括结合酶对叶酰多-γ-谷氨酸底物的水解、L-谷氨酸脱氢酶将谷氨酸转化为α-酮戊二酸以及比色法测量α-酮戊二酸的BDBH衍生物。通过该方法可以测量从底物中释放出的低至1 nmol的谷氨酸，这非常适合用于各种结合酶制剂的测定。此外，该方法应为各种叶酸和抗叶酸多聚谷氨酸的酶促水解提供一种通用的测定方法。
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