Dahlmann B, Kuehn L, Heinrich P C, Kirschke H, Wiederanders B
Biochemische Abteilung, Diabetes Forschungsinstitut, Düsseldorf, F.R.G.
Biochim Biophys Acta. 1989 May 31;991(2):253-62. doi: 10.1016/0304-4165(89)90113-x.
From rat skeletal muscle tissue we have isolated and purified a proteolytic activity of molecular mass 750 kDa. The enzyme, designated 'proteinase I', which has been found to be located in capillaries of skeletal muscle tissue, catalyzes the hydrolysis of Z-Phe-Arg-MCA and [14C]methylcasein and this process is activated about 2-fold by ATP. As judged by SDS-polyacrylamide gel electrophoresis the subunit pattern of 'proteinase I' is similar to alpha-macroglobulin. Immunoelectrophoretic analyses of 'proteinase I' with antisera to rat alpha 1-macroglobulin, alpha 2-macroglobulin, and rat liver cathepsins reveal that this high-molecular-mass proteinase is a complex of alpha 1-macroglobulin and the cysteine proteinases cathepsin B, H and L. A similar 'proteinase' has been isolated from rat serum. Two ATP-activated high molecular-mass proteinases that have been previously identified in liver and heart muscle by other investigators equally show a positive immunological reaction with the antiserum raised against 'proteinase I'. From these data, together with results presented in an accompanying paper (Kuehn, L., Dahlmann, B., Gauthier, F. and Neubauer, H.-P. (1989) Biochim. Biophys. Acta 991, 263), we conclude that the ATP-stimulated high-molecular-mass proteolytic activity is partly due to the presence of a complex of alpha-macroglobulin and cysteine proteinases.
我们从大鼠骨骼肌组织中分离并纯化出了一种分子量为750 kDa的蛋白水解活性物质。这种酶被命名为“蛋白酶I”，已发现其位于骨骼肌组织的毛细血管中，它能催化Z-苯丙氨酸-精氨酸-甲基香豆素酰胺（Z-Phe-Arg-MCA）和[14C]甲基酪蛋白的水解，且该过程可被ATP激活约2倍。通过SDS-聚丙烯酰胺凝胶电泳判断，“蛋白酶I”的亚基模式与α-巨球蛋白相似。用针对大鼠α1-巨球蛋白、α2-巨球蛋白和大鼠肝脏组织蛋白酶的抗血清对“蛋白酶I”进行免疫电泳分析表明，这种高分子量蛋白酶是α1-巨球蛋白与半胱氨酸蛋白酶组织蛋白酶B、H和L的复合物。已从大鼠血清中分离出一种类似的“蛋白酶”。其他研究人员先前在肝脏和心肌中鉴定出的两种ATP激活的高分子量蛋白酶，同样与针对“蛋白酶I”产生的抗血清呈现阳性免疫反应。根据这些数据以及随附论文（Kuehn, L., Dahlmann, B., Gauthier, F. 和Neubauer, H.-P. (1989) Biochim. Biophys. Acta 991, 263）中给出的结果，我们得出结论，ATP刺激的高分子量蛋白水解活性部分归因于α-巨球蛋白和半胱氨酸蛋白酶复合物的存在。
