Monsees T, Geiger R, Miska W
Department of Dermatology and Andrology, Justus Liebig University, Giessen, Germany.
J Biolumin Chemilumin. 1995 Jul-Aug;10(4):213-8. doi: 10.1002/bio.1170100404.
The use of 6-(N-acetyl-L-phenylalanyl)-aminoluciferin as a novel substrate for alpha-chymotrypsin has been demonstrated. The kinetic parameters determined are KM = 0.38 mmol/L, kcat = 6.5 s-1 and kcat/kM = 17,100 (L/mol s). The test principle of the coupled assay is the release of aminoluciferin by enzymatic cleavage of 6-(N-acetyl-L-phenylalanyl)-aminoluciferin. Aminoluciferin is oxidized, with light emission, by firefly luciferase (Photinus pyralis) and can be quantified in a luminometric assay. The detection limit for chymotrypsin was found to be 0.3 ng per assay. 6-(N-acetyl-L-phenylalanyl)-aminoluciferin has been synthesized as an example for a new class of highly sensitive substrates. By modification of the peptide residue these new substrates may be suitable for ultrasensitive detection of different proteinases.
已证明6-(N-乙酰-L-苯丙氨酰基)-氨基荧光素可作为α-胰凝乳蛋白酶的新型底物。所测定的动力学参数为：KM = 0.38 mmol/L，kcat = 6.5 s-1，kcat/kM = 17,100 (L/mol·s)。偶联测定的测试原理是通过酶促裂解6-(N-乙酰-L-苯丙氨酰基)-氨基荧光素释放氨基荧光素。氨基荧光素被萤火虫荧光素酶（Photinus pyralis）氧化并发光，可在发光测定中进行定量。发现胰凝乳蛋白酶的检测限为每次测定0.3 ng。已合成6-(N-乙酰-L-苯丙氨酰基)-氨基荧光素作为一类新型高灵敏度底物的示例。通过修饰肽残基，这些新底物可能适用于不同蛋白酶的超灵敏检测。
