Physicochemical Characterization of A and B Subunits of Shiga Toxin and Reconstitution of Holotoxin from Isolated Subunits

• Published in: MICROBIOLOGY and IMMUNOLOGY Vol. 31; no. 3; pp. 189 - 197
• Main Authors: Yutsudo, Takashi, Honda, Takeshi, Miwatani, Toshio, Takeda, Yoshifumi
• Format: Journal Article
• Language: English
• Published: Australia Blackwell Publishing Ltd 1987; Center For Academic Publications Japan
• Subjects: Amino Acids - analysis; Animals; Bacterial Toxins - analysis; Bacterial Toxins - isolation & purification; Bacterial Toxins - toxicity; Chromatography, High Pressure Liquid; Electrophoresis, Polyacrylamide Gel; Isoelectric Point; Mice; Shiga Toxins; Shigella dysenteriae; Trypsin
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1987.tb03083.x

The A and B subunits of Shiga toxin were isolated by high performance liquid chromatography and their physicochemical properties were examined. The A subunit of Shiga toxin purified from culture supernatant was not nicked, but it could be nicked in vitro by trypsin. The isoelectric points of the A and B subunits were determined to be 8.2 and 5.8, respectively. Amino acid compositions of the two subunits were also determined. The isolated A and B subunits were reconstituted to form active holotoxin which showed lethal activity to mice which was similar to that of native Shiga toxin.