Tonnesen M G, Klempner M S, Austen K F, Wintroub B U
J Clin Invest. 1982 Jan;69(1):25-30. doi: 10.1172/jci110437.
A human neutrophil protease, initially termed neutral peptide-generating protease, has been shown to cleave angiotensin II directly from angiotensinogen and has been identified as leukocyte cathepsin G. When purified neutrophils were disrupted by nitrogen cavitation and fractionated by differential centrifugation, 44 and 24% of the angiotensin II-generating activity was in the lysosomal and undisrupted cell fractions, respectively. Cytochalasin B-treated human neutrophils stimulated with N-formyl-L-methionyl-L-leucyl-L-phenylalanine released beta-glucuronidase, lysozyme, and angiotensin II-generating protease in a dose-dependent fashion, consistent with localization of this protease to the neutrophil granule. Individually purified angiotensin II-generating protease and cathepsin G had similar proteolytic and esterolytic activity for angiotensinogen and N-benzoyl-L-tyrosine ethyl ester on a weight basis, exhibited identical mobilities by SDS-gradient polyacrylamide gel electrophoresis and pH 4.3 disc-gel electrophoresis, and gave precipitin lines of antigenic identity on Ouchterlony analysis with goat antibody to the angiotensin II-generating protease. Thus, the angiotensin II-generating protease of human neutrophils has been identified as cathepsin G on the basis of subcellular localization, substrate specificity, physicochemical characteristics, and antigenic identity.
一种最初被称为中性肽生成蛋白酶的人类中性粒细胞蛋白酶，已被证明可直接从血管紧张素原中裂解出血管紧张素II，并已被鉴定为白细胞组织蛋白酶G。当通过氮空化破坏纯化的中性粒细胞并通过差速离心进行分级分离时，分别有44%和24%的血管紧张素II生成活性存在于溶酶体和未破坏的细胞级分中。用N-甲酰-L-蛋氨酰-L-亮氨酰-L-苯丙氨酸刺激经细胞松弛素B处理的人类中性粒细胞，会以剂量依赖的方式释放β-葡萄糖醛酸酶、溶菌酶和血管紧张素II生成蛋白酶，这与该蛋白酶定位于中性粒细胞颗粒一致。单独纯化的血管紧张素II生成蛋白酶和组织蛋白酶G在重量基础上对血管紧张素原和N-苯甲酰-L-酪氨酸乙酯具有相似的蛋白水解和酯水解活性，通过SDS梯度聚丙烯酰胺凝胶电泳和pH 4.3圆盘凝胶电泳显示出相同的迁移率，并且在用针对血管紧张素II生成蛋白酶的山羊抗体进行免疫双扩散分析时产生抗原同一性的沉淀线。因此，基于亚细胞定位、底物特异性、物理化学特性和抗原同一性，人类中性粒细胞的血管紧张素II生成蛋白酶已被鉴定为组织蛋白酶G。
