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Isolation ofEscherichia coli heat-labile enterotoxin by affinity chromatography: Characterization of subunits

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Abstract

This paper describes the isolation ofEscherichia coli heat-labile enterotoxin (LT) by affinity chromatography on an anti-cholera toxin immunoglobulin-Sepharose column, and the subunit composition of crude and affinity-isolated LT. LT and its subunits were assayed with ganglioside (GM1)-ELISA, immunodiffusion, skin toxicity, and broken cell adenylate cyclase activation methods. The results show that the immunoaffinity method, applied to LT of different strains and batches, yielded about 100-fold purification with approximately 50% recovery of LT antigen. LT was shown to contain a GM1-ganglioside binding subunit as well as another subunit which does not bind to GM1 but activates adenylate cyclase. Immunodiffusion tests showed that the two LT subunits were immunologically related to but not identical with, respectively, the B and A subunits of cholera toxin. The LT “A” and “B” subunits were present in similar proportions in the affinity-isolated and crude LT preparations, but in the purified fraction they had only partially reassociated into holotoxin.

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Authors and Affiliations

  1. State of Israel Laboratory of Poultry Diseases, Joffa Street 191, P. O. B. 13088, Jerusalem, Israel

    Moshe Wolk

  2. Insitute of Medical Microbiology, University of Göteborg, Guldhedsgatan 10, S-41346, Göteborg, Sweden

    Ann-Mari Svennerholm & Jan Holmgren

Authors
  1. Moshe Wolk
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  2. Ann-Mari Svennerholm
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  3. Jan Holmgren
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Wolk, M., Svennerholm, AM. & Holmgren, J. Isolation ofEscherichia coli heat-labile enterotoxin by affinity chromatography: Characterization of subunits. Current Microbiology 3, 339–344 (1980). https://doi.org/10.1007/BF02601898

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