North M J, Harwood J M
Biochim Biophys Acta. 1979 Jan 12;566(1):222-33. doi: 10.1016/0005-2744(79)90264-x.
Proteinase activity in the cellular slime mould Dictyostelium discoideum has been analyzed by electrophoresis on polyacrylamide gels containing denatured hemoglobin. At least eight bands due to acid proteinases have been defined using extracts of myxamoebae, four bands A-D which move faster than the fifth and major band E, a minor band E' which moves just behind E and two slow bands G and H. Fruiting body formation was accompanied by the appearance of one new proteinase band F. The proteinases were present in extracts of both axenically-grown and bacterially-grown cells. Differences between the pH dependence and stability of the individual proteinases were detected. Inhibitor studies suggested that the faster proteinases A-D may be cathepsin B-like, whilst the slower enzymes E, E' and F do not fit readily into any known group of proteinases since they were sensitive to HgCl2 but not to other inhibitors of cathepsin B and not to inhibitors of cathepsin D-like proteinases under standard conditions. None of the proteinases was apparently formed during or after preparation of extracts and the proteinases could be re-run on polyacrylamide gels to give only the band expected from the first run. The bands are believed to reflect multiple proteinase activities within the cell.
通过在含有变性血红蛋白的聚丙烯酰胺凝胶上进行电泳,分析了细胞黏菌盘基网柄菌中的蛋白酶活性。使用变形虫提取物已确定了至少八条由酸性蛋白酶产生的条带,四条条带A - D比第五条也是主要的条带E移动得更快,一条次要条带E'紧挨着E后面移动,还有两条慢速条带G和H。子实体形成伴随着一条新的蛋白酶条带F的出现。蛋白酶存在于无菌培养和细菌培养细胞的提取物中。检测到了各个蛋白酶在pH依赖性和稳定性方面的差异。抑制剂研究表明,较快的蛋白酶A - D可能类似于组织蛋白酶B,而较慢的酶E、E'和F不太容易归入任何已知的蛋白酶类别,因为在标准条件下它们对HgCl2敏感,但对组织蛋白酶B的其他抑制剂以及组织蛋白酶D样蛋白酶的抑制剂不敏感。在提取物制备过程中或之后显然没有形成任何蛋白酶,并且这些蛋白酶可以在聚丙烯酰胺凝胶上重新进行电泳,结果只会出现第一次电泳预期的条带。据信这些条带反映了细胞内多种蛋白酶的活性。