Preparation and characterization of an active fragment of colicin E3.

Abstract

The conditions of digestion of colicin E3 with trypsin were examined to obtain an active fragment (T2A) of colicin E3, and a method suitable for large-scale preparation of T2A was developed. The T2A preparation thus obtained was homogeneous on SDS-polyacrylamide gel electrophoresis and the molecular weight was estimated to be about 11,000. T2A was composed of 97 amino acid residues and was rich in basic amino acids; methionine, valine, cysteine, and cystine were absent. The N-terminal residue was lysine and the structure near the C-terminus was -Lys-Lys-Tyr-Leu. Since T2A had no lysine or arginine residue at the C-terminus and since the C-terminal structure was identical to that of protein A, it was concluded that T2A was derived from the C-terminal region of protein A. No clear differences were detected among T2A preparations obtained from 3 different fractions of colicin E3, suggesting that the apparent homogeneity of colicin E3 does not involve the T2A region.