Isolation and some properties of A and B subunits of Vero toxin 2 and in vitro formation of hybrid toxins between subunits of Vero toxin 1 and Vero toxin 2 from Escherichia coli O157:H7

Abstract

Purified Vero toxin 2 (VT2) was separated into A and B subunits by treatment with 6 M urea in 0.1 M propionic acid (pH 4.0). The isoelectric points of the isolated A and B subunits were determined to be 8.1 and 4.1, respectively. The A subunit of the purified VT2 was not nicked, but could be nicked in vitro by trypsin. Biologically active toxin was reconstituted from the isolated A and B subunits of VT2. Hybrid toxins with biological activity were obtained in vitro from the A subunit of Vero toxin 1 (VT1) and the B subunit of VT2, and from the A subunit of VT2 and the B subunit of VT1. The hybrid toxins showed similar cytotoxicity to native VT1 and VT2 on Vero cells. The in vitro formations of hybrid toxins were confirmed by polyacrylamide disc gel electrophoresis.