“In vitro” Study of Basement Membrane Degradation by the Cysteine Proteinases, Cathepsins B, B-Like and L. Digestion of Collagen IV, Laminin, Fibronectin, and Release of Gelatinase Activities front Basement Membrane Fibronectin

Abstract

We have studied the soluble fragments obtained from bovine lens capsules after digestion by the cysteine proteinases cathepsins B, B-like and L. These proteinases liberated collagen IV, laminin and fibronectin fragments, as shown by immunoblotting. Sodium dodecyl sulfate treatment of digested capsules gave a soluble material used for subsequent fractionation and immunochemical study. Comparison of both results demonstrate the ability of these cathepsins to degrade a basement membrane at near neutral pH values. The differences observed in the size and the number of fragments suggest that the three proteinases exhibit similar specificities in basement membrane digestion, as shown previously. Nevertheless, cathepsin L seems to be more effective than cathepsins B and B-like. From this study, cysteine proteinases could be associated to basement membrane destruction. Soluble cysteine proteinase digests of bovine lens capsules showed several bands of gelatinolytic activity by gelatin zymography. Three major bands of 77, 60 and 45 kDa were seen whatever the cysteine proteinase used. These bands were identified as fibronectin fragments. Thus cysteine proteinases can activate the latent proteinase fibronectin from basement membrane leading to a new "metastatic cascade". This would be an important factor in the "in vivo" basement membrane dissolution observed during tumor invasion.