Fragmentation of colicins A and E1 by cell surface proteases.

Interaction of either colicin A or E1 with the surface of Escherichia coli cells resulted in extensive cleavage of the colicins into many peptide fragments in the molecular weight range of 10,000 to 30,000 released into the supernatants of colicin-cell mixtures. The protease inhibitor P-aminobenzamidine inhibited the cleavage of colicin A and enhanced colicin killing activity, suggesting that proteolysis is not required for the killing action of colicin. Fragments derived from the supernatants of the mixtures were inactive against sensitive cells. Proteolytic activity against both colicins was localized primarily in the outer membrane fraction of the cell envelope. At least two distinct protease activities appear to be present. Examination of the patterns of cleavage and inactivation of the colicins by a series of resistant mutants indicates that specific colicin receptors play no essential role in colicin proteolysis. In addition, evidence is presented that adsorption of colicin to specific receptors is a reversible process.