Physicochemical characterization of A and B subunits of Shiga toxin and reconstitution of holotoxin from isolated subunits.
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.
Yutsudo
T
T
Honda
T
T
Miwatani
T
T
Takeda
Y
Y
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis* / Q000302:isolation & purification / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D007526:Isoelectric Point; D051379:Mice; D022603:Shiga Toxins; D012762:Shigella dysenteriae; D014357:Trypsin
D000596:Amino Acids; D001427:Bacterial Toxins; D022603:Shiga Toxins; D014357:Trypsin