Selective inhibition of binding of Bacillus thuringiensis Cry1Ab toxin to cadherin-like and aminopeptidase proteins in brush border membranes and dissociated epithelial cells from Bombyx mori

M Sales Ibiza-Palacios , Juan Ferré , Satoshi Higurashi , Kazuhisa Miyamoto , Ryoichi Sato , Baltasar Escriche (1)

1 Dep. Genética

Binding analyses with denatured epithelial membrane proteins from Bacillus thuringiensis (Bt) evidenced at least two kinds of proteins, aminopeptidases N (APNs) and cadherin-ike proteins, as possible receptors for the Cry1A class of Bt toxins. Two alternative models have been proposed, both based on initial toxin binding to a cadherin-like protein, but one involving APN and the other not. We have used two Bombyx mori strains (J65 and Kin), which are highly susceptible to Cry1Ab, to study the role of these two types of receptors on Cry1Ab toxin binding and cytotoxicity by means of the inhibitory effect of antibodies. Brush border membrane vesicles (BBMV) of strain J65 incubated with labeled 125}I-Cry1Ab revealed a marked reduction in reversible and irreversible binding when anti-BtR175 (a cadherin-like protein) was used for BBMV pretreatment. By contrast, the anti-APN1 antibody specifically affected the irreversible binding, while the reversible binding component was not affected. This is the first time that binding of Cry1Ab to APN1 and to a cadherin-like protein from BBMV in solution has been shown. Dissociated epithelial cells from the Kin strain were used to test the inhibitory effect of the antibodies on the cytotoxicity of Cry1Ab. Preincubation of the cells with the anti-BtR175 antibody conferred protection against Cry1Ab, but not the anti-PN1 antibody. Therefore, our results seem to support the two models of the mode of action of Cry1Ab in Lepidoptera, depending on whether BBMV or intact dissociated cells are used, suggesting that both pathways may co-operate for the toxicity of Cry1A toxins in vivo

Mots clés: Life Sciences

M Sales Ibiza-Palacios, Juan Ferré, Satoshi Higurashi, Kazuhisa Miyamoto, Ryoichi Sato, et al.. Selective inhibition of binding of Bacillus thuringiensis Cry1Ab toxin to cadherin-like and aminopeptidase proteins in brush border membranes and dissociated epithelial cells from Bombyx mori. Biochemical Journal, 2007, 409 (1), pp.215-221. ⟨10.1042/BJ20070956⟩. ⟨hal-00478848⟩