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. 1980 Oct;30(1):170–179. doi: 10.1128/iai.30.1.170-179.1980

Characterization of Shigella dysenteriae 1 (Shiga) toxin purified by anti-Shiga toxin affinity chromatography.

A D O'Brien, G D LaVeck, D E Griffin, M R Thompson
PMCID: PMC551292  PMID: 7002787

Abstract

Shigella dysenteriae 1 (Shiga) toxin was purified from whole-cell lysates by antitoxin affinity column chromatography, radioiodination, and Sephacryl S-200 gel filtration of 125I-labeled affinity column eluates. Two chromatographic peaks were observed. The percentage of radioactivity in peak I samples immunoprecipitated with antitoxin ranged from 95 to 100%. A pool of samples from this first peak contained over 90% of the HeLa-cell-cytotoxic units applied to the column and was enterotoxic for rabbit ileal loops and lethal for rabbits. This radiolabeled material migrated as a single cytotoxic band after nondenaturing polyacrylamide gel electrophoresis, but formed three bands, of 33,000, 29,000, and 4,000 to 7,000 daltons, after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In addition, material estimated as 7,000 daltons by Bio-Gel P-10 chromatography could be generated by treatment of S-200 peak I samples with 8 M urea. Pooled fractions from the second S-200 peak were separable into several low-molecular-weight peaks on a P-10 column. One of these P-10 peaks (7,000 daltons) was 27% immunoprecipitable with antitoxin. These data indicate that three of the known biological activities of Shiga toxin are associated with a 33,000-dalton substance which can be dissociated into 29,000- and 4,000- to 7,000-dalton components.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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