Guinec N, Pagano M, Dalet-Fumeron V, Engler R
Laboratoire de Biochimie, Faculté de Médecine Broussais Hôtel Dieu, Paris, France.
Biol Chem Hoppe Seyler. 1990 May;371 Suppl:239-54.
We have examined the biochemical degradation of an isolated basement membrane matrix (bovine lens capsule) by human liver cathepsins B, H and L and the cathepsin B-like proteinase from malignant ascitic fluid. This study was carried out using two different methods: The first strategy was to follow the liberation of soluble proteins and peptides as a function of time at different pHs. Then the digestion products were characterized, as collagen IV, fibronectin and laminin fragments, using monospecific polyclonal antibodies and a quantitative dot-blot analysis. From these results, the ability of the four proteinases to digest "in vitro" intact bovine lens capsule in the physiological pH range is demonstrated. Cathepsin L is the most powerful against the three membrane components studied. As shown by electroelution and immunochemical quantitation, the digestion would be a consequence of proteinases binding to the capsule. With intact basement membrane as a substrate a "in vitro" molecular analysis of this digestion process was possible by these methods. On this basis, the "in vivo" secretion of cysteine proteinases during malignancy would be related to the local basement membrane dissolution associated with tumor invasion.
我们研究了人肝脏组织蛋白酶B、H和L以及来自恶性腹水的类组织蛋白酶B蛋白酶对分离的基底膜基质（牛晶状体囊）的生化降解作用。本研究采用了两种不同的方法：第一种策略是在不同pH值下，跟踪可溶性蛋白质和肽随时间的释放情况。然后，使用单特异性多克隆抗体和定量斑点印迹分析，将消化产物鉴定为IV型胶原、纤连蛋白和层粘连蛋白片段。从这些结果可以证明，这四种蛋白酶在生理pH范围内“体外”消化完整牛晶状体囊的能力。组织蛋白酶L对所研究的三种膜成分的作用最强。通过电洗脱和免疫化学定量分析表明，消化是蛋白酶与囊结合的结果。通过这些方法，以完整的基底膜为底物，对该消化过程进行“体外”分子分析成为可能。在此基础上，恶性肿瘤期间半胱氨酸蛋白酶的“体内”分泌可能与肿瘤侵袭相关的局部基底膜溶解有关。
