Sogawa K, Takahashi K
J Biochem. 1976 Dec;80(6):1443-6. doi: 10.1093/oxfordjournals.jbchem.a131419.
A novel neutral protease(s), which is presumably membrane-bound, was found in monkey liver using heat-denatured casein as a substrate and was separated from other major catheptic proteases by successive procedures of gel filtration on Ultrogel AcA 22, solubilization by deoxycholate and gel filtration on Sepharose 6B. The enzyme(s) showed maximal activity at pH 8.0, and was strongly inhibited by DFP and PMSF. Many other reagents tested, including TPCK, TLCK, pCMB, iodoacetic acid, and EDTA, were without marked effect on the activity. Activation of the enzyme(s) by NaCl was not observed.
利用热变性酪蛋白作为底物，在猴肝脏中发现了一种可能与膜结合的新型中性蛋白酶，通过在Ultrogel AcA 22上进行连续的凝胶过滤、用脱氧胆酸盐增溶以及在Sepharose 6B上进行凝胶过滤等步骤，将其与其他主要的组织蛋白酶分离。该酶在pH 8.0时表现出最大活性，并受到二异丙基氟磷酸（DFP）和苯甲基磺酰氟（PMSF）的强烈抑制。所测试的许多其他试剂，包括对甲苯磺酰-L-苯丙氨酸氯甲基酮（TPCK）、对甲苯磺酰-L-赖氨酸氯甲基酮（TLCK）、对氯汞苯甲酸（pCMB）、碘乙酸和乙二胺四乙酸（EDTA），对其活性没有明显影响。未观察到氯化钠对该酶的激活作用。
