Characterization of purified Shiga toxin from Shigella dysenteriae 1.
Shiga toxin was purified from the culture supernatant of Shigella dysenteriae 1 by ammonium sulfate fractionation, DEAE-cellulose column chromatography and repeated chromatofocusing column chromatography. About 1.6 mg of purified Shiga toxin was obtained from 15 liters of culture with a yield of about 27%. The molecular weight of purified Shiga toxin was estimated to be 62,000. The toxin consisted of A and B subunits with molecular weights of about 30,000 and 5,000-6,000, respectively. The isoelectric point of purified Shiga toxin was 7.0. Purified Shiga toxin showed the following biological activities: lethal toxicity to mice when injected intraperitoneally with an LD50 of 28 ng per mouse; cytotoxicity to Vero cells, killing about 50% of the cells at 1 pg and all of the cells at 10 pg; and fluid accumulation in rabbit ileal loops at a concentration of more than 1 microgram.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis* / Q000302:isolation & purification; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002848:Chromatography, DEAE-Cellulose; D000882:Haplorhini; D007082:Ileum / Q000187:drug effects; D007525:Isoelectric Focusing; D007928:Lethal Dose 50; D051379:Mice; D008970:Molecular Weight; D011817:Rabbits; D022603:Shiga Toxins; D012762:Shigella dysenteriae / Q000032:analysis*
D000596:Amino Acids; D001427:Bacterial Toxins; D022603:Shiga Toxins