Estimation of the radius of the pores formed by the Bacillus thuringiensis Cry1C delta-endotoxin in planar lipid bilayers.
Pore formation constitutes a key step in the mode of action of Bacillus thuringiensis delta-endotoxins and various activated Cry toxins have been shown to form ionic channels in receptor-free planar lipid bilayers at high concentrations. Multiple conductance levels have been observed with several toxins, suggesting that the channels result from the multimeric assembly of a variable number of toxin molecules. To test this possibility, the size of the channels formed by Cry1C was estimated with the non-electrolyte exclusion technique and polyethylene glycols of various molecular weights. In symmetrical 300 mM KCl solutions, Cry1C induced channel activity with 15 distinct conductance levels ranging from 21 to 246 pS and distributed in two main conductance populations. Both the smallest and largest conductance levels and the mean conductance values of both populations were systematically reduced in the presence of polyethylene glycols with hydrated radii of up to 1.05 nm, indicating that these solutes can penetrate the pores formed by the toxin. Larger polyethylene glycols had little effect on the conductance levels, indicating that they were excluded from the pores. Our results indicate that Cry1C forms clusters composed of a variable number of channels having a similar pore radius of between 1.0 and 1.3 nm and gating synchronously.
Peyronnet
Olivier
O
Groupe de recherche en transport membranaire, Université de Montréal, P.O. Box 6128, Centre Ville Station, Montreal, Quebec, Canada H3C 3J7.
Nieman
Brian
B
Généreux
Francis
F
Vachon
Vincent
V
Laprade
Raynald
R
Schwartz
Jean-Louis
JL
D001413:Bacillus thuringiensis / Q000502:physiology*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000502:physiology*; D001427:Bacterial Toxins; D004731:Endotoxins / Q000502:physiology*; D006460:Hemolysin Proteins; D007473:Ion Channels / Q000502:physiology; D008051:Lipid Bilayers / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D011092:Polyethylene Glycols