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Lepidopteran-specific crystal toxins from Bacillus thuringiensis form cation- and anion-selective channels in planar lipid bilayers

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Summary

Previous studies in our laboratory have shown that CryIC, a lepidopteran-specific toxin from Bacillus thuringiensis, triggers calcium and chloride channel activity in SF-9 cells (Spodoptera frugiperda, fall armyworm). Chloride currents were also observed in SF-9 membrane patches upon addition of CryIC toxin to the cytoplasmic side of the membrane. In the present study the ability of activated CryIC toxin to form channels was investigated in a receptor-free, artificial phospholipid membrane system. We demonstrate that this toxin can partition in planar lipid bilayers and form ion-selective channels with a large range of conductances. These channels display complex activity patterns, often possess subconducting states and are selective to either anions or cations. These properties appeared to be pH dependent. At pH 9.5, cation-selective channels of 100 to 200 pS were most frequently observed. Among the channels recorded at pH 6.0, a 25–35 pS anion-selective channel was often seen at pH 6.0, with permeation and kinetic properties similar to those of the channels previously observed in cultured lepidopteran cells under comparable pH environment and for the same CryIC toxin doses. We conclude that insertion of CryIC toxin in SF-9 cell native membranes and in artificial planar phospholipid bilayers may result from an identical lipid-protein interaction mechanism.

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Authors and Affiliations

  1. Biotechnology Research Institute, National Research Council, H4P 2R2, Montreal, Quebec, Canada

    Jean-Louis Schwartz, Line Garneau, Luke Masson & Roland Brousseau

  2. Membrane Transport Research Group, University of Montreal, H3C 3J7, Montreal, Quebec, Canada

    Diane Savaria

  3. Department of Physiology and Biophysics, Faculty of Medicine, University of Sherbrooke, J1H 5N4, Sherbrooke, Quebec, Canada

    Eric Rousseau

Authors
  1. Jean-Louis Schwartz
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  2. Line Garneau
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  3. Diane Savaria
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  4. Luke Masson
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  5. Roland Brousseau
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  6. Eric Rousseau
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Additional information

The assistance of A. Mazza and G.A.R. Mealing is gratefully acknowledged. The trypsin-activated, HPLC-purified CryIC toxin isolated from B. thuringiensis var. entomocidus crystal was a kind gift from M. Pusztai, Institute for Biological Sciences, NRC, Ottawa.

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Schwartz, JL., Garneau, L., Savaria, D. et al. Lepidopteran-specific crystal toxins from Bacillus thuringiensis form cation- and anion-selective channels in planar lipid bilayers. J. Membarin Biol. 132, 53–62 (1993). https://doi.org/10.1007/BF00233051

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  • DOI: https://doi.org/10.1007/BF00233051

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