Rindler-Ludwig R, Braunsteiner H
Biochim Biophys Acta. 1975 Feb 27;379(2):606-17. doi: 10.1016/0005-2795(75)90167-1.
Three cationic proteins from the granules of human neutrophil granulocytes were obtained in a high degree of purity be means of affinity chromatography on 4-phenylbutylamine-Sepharose. Together with lysozyme, the three cationic proteins exhibit the highest electrophoretic mobility toward the cathode in acrylamide gels at moderately acid pH, among the granule constituents that are solubilized in 0.1 M phosphate buffer, pH 7.0, containing 1 M NaCl. The three cationic proteins represent a group of "neutral proteases" distinct from elastase and collagenase. They hydrolyze casein, azocasein and the chymotrypsin substrate N-acetyl-L-tyrosine ethyl ester. Optimal activity is found at pH 7.4-7;5. The enzymes are inhibited by the specific chymotrypsin inhibitor N-tosyl-L-phenylalanylchloromethane and by the naturally occurring inhibitors alpha-antichymotrypsin, alpha-1-antitrypsin, as well as by the trypsin inhibitors from soy beans and limabeans.
通过在4-苯基丁胺-琼脂糖上进行亲和层析，从人中性粒细胞的颗粒中获得了三种纯度很高的阳离子蛋白。在0.1M pH 7.0的磷酸盐缓冲液（含1M氯化钠）中溶解的颗粒成分中，这三种阳离子蛋白与溶菌酶一起，在中等酸性pH条件下的丙烯酰胺凝胶中，向阴极表现出最高的电泳迁移率。这三种阳离子蛋白代表了一组不同于弹性蛋白酶和胶原酶的“中性蛋白酶”。它们能水解酪蛋白、偶氮酪蛋白和胰凝乳蛋白酶底物N-乙酰-L-酪氨酸乙酯。在pH 7.4 - 7.5时发现最佳活性。这些酶受到特异性胰凝乳蛋白酶抑制剂N-甲苯磺酰-L-苯丙氨酰氯甲烷、天然存在的抑制剂α-抗胰凝乳蛋白酶、α-1-抗胰蛋白酶以及来自大豆和利马豆的胰蛋白酶抑制剂的抑制。
