Purification and molecular properties of a new colicin

The process of isolation and purification of a new colicin isolated from a Citrobacter strain is described. Escherichia coli sensitive cells are protected by vitamin B12 from the action of this bacteriocin; this suggests that it belongs to the E group of colicins. Therefore, we have called it colicin E4. It has a molecular weight of 56 000 and two molecular forms of isoelectric points 9.4 and 8.2 are separated in electrofocusing on polyacrylamide gels. It has a sedimentation coefficient of 3.4 S and the absorption coefficient A1(280%) nm is 6.23 cm(-1). Using an antibody raised against pure colicin E4, no cross-reaction was detected against colicins A, E1 or K. The physiological effect of colicin E4 on sensitive cells is very similar to that of colicins E1, K or I which disrupt the energized membrane state.