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Cell surface-associated and released proteolytic activities of bovine aorta endothelial cells.

We have demonstrated cell surface-associated and released proteolytic activity on bovine aorta endothelial cells, representing normal cells with regulated invasive properties. To demonstrate these proteolytic activities on viable cells grown in monolayer cultures, a new method was developed. The method consists of rolling modified plastic beads carrying covalently-linked [125I]-labeled casein in contact with the cell surfaces or adjacent to the cell monolayers without actual contact. The rate of radioactive peptide release is proportional to the proteolytic activity. Released proteases are detected when no contact occurs between the substrate and the cells. When the bead-substrate complex is rolled over the surface of endothelial cells, a significant increase in released labeled peptides is observed which represents a cell surface-associated proteolytic activity. These activities may be relevant to the endothelial cell's invasive capacity and appear to be similar to the neutral proteolytic activity of transformed cells.

Animals↗

Calcium-induced cleavage and breakdown of spectrin in the rat lens.

Incubation of intact rat lenses under conditions that stimulated a net influx of calcium resulted in a pronounced loss of transparency and a major decrease in the levels of spectrin. The progressive loss of this cytoskeletal component coincided with the appearance of polypeptides of approximately 150 kDa which showed immunoreactivity with an antibody raised to spectrin. These bands disappeared on further incubation. It is, therefore, suggested that a calcium-activated protease is present in the lens which is capable of degrading spectrin by the initial removal of approximately 90 kDa fragments. This process calcium-induced proteolysis may be the basis for the cytoskeletal reorganisation observed during the differentiation of lens fibre cells and may be involved in cataract development.

Animals↗