Rush R S, Mitas M, Powers J C, Tanaka T, Hersh L B
Arch Biochem Biophys. 1984 Jun;231(2):390-9. doi: 10.1016/0003-9861(84)90402-8.
Rat brain neutral endopeptidase ("Enkephalinase") was shown to hydrolyze a series of fluorogenic substrates of the general structure 2-aminobenzoyl-(amino acid)n- leucylalanylglycine -4- nitrobenzylamide . The hydrolysis of these substrates was competitively inhibited by Leu5-enkephalin, demonstrating that these are indeed substrates for the rat brain neutral endopeptidase. Cleavage of the fluorogenic substrates yielded leucylalanylglycine -4- nitrobenzylamide as a common product. In addition, a series of inhibitors previously shown to inhibit thermolysin-like enzymes inhibited the hydrolysis of both Leu5-enkephalin and the synthetic substrates. The results of this study (a) demonstrate that the enkephalin-degrading endopeptidase is similar in specificity to thermolysin, (b) provide a continuous sensitive assay system for the enzyme, and (c) point out the potential use of this substrate class for probing the specificity of the enzyme.
大鼠脑中性内肽酶（“脑啡肽酶”）被证明能水解一系列具有通式2-氨基苯甲酰-（氨基酸）n-亮氨酰丙氨酰甘氨酸-4-硝基苄酰胺的荧光底物。这些底物的水解受到亮氨酸脑啡肽的竞争性抑制，表明它们确实是大鼠脑中性内肽酶的底物。荧光底物的裂解产生亮氨酰丙氨酰甘氨酸-4-硝基苄酰胺作为共同产物。此外，先前显示能抑制嗜热菌蛋白酶样酶的一系列抑制剂抑制了亮氨酸脑啡肽和合成底物的水解。本研究结果（a）表明，降解脑啡肽的内肽酶在特异性上与嗜热菌蛋白酶相似，（b）为该酶提供了一个连续灵敏的检测系统，（c）指出了这类底物在探究酶特异性方面的潜在用途。
