Binding analyses of Cry1Ab and Cry1Ac with membrane vesicles from Bacillus thuringiensis-resistant and -susceptible Ostrinia nubilalis

Biochem Biophys Res Commun. 2004 Oct 8;323(1):52-7. doi: 10.1016/j.bbrc.2004.08.054.


The binding properties of Bacillus thuringiensis toxins to brush border membrane vesicles of Dipel-resistant and -susceptible Ostrinia nubilalis larvae were compared using ligand-toxin immunoblot analysis, surface plasmon resonance (SPR), and radiolabeled toxin binding assays. In ligand-toxin immunoblot analysis, the number of Cry1Ab or Cry1Ac toxin binding proteins and the relative toxin binding intensity were similar in vesicles from resistant and susceptible larvae. Surface plasmon resonance with immobilized activated Cry1Ab toxin indicated that there were no significant differences in binding with fluid-phase vesicles from resistant and susceptible larvae. Homologous competition assays with radiolabeled Cry1Ab and Cry1Ac toxin and vesicles from resistant and susceptible larvae resulted in similar toxin dissociation constants and binding site concentrations. Heterologous competition binding assays indicated that Cry1Ab and Cry1Ac completely competed for binding, thus they share binding sites in the epithelium of the larval midguts of O. nubilalis. Overall, the binding analyses indicate that resistance to Cry1Ab and Cry1Ac in this Bt-resistant strain of O. nubilalis is not associated with a loss of toxin binding.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Bacillus thuringiensis / metabolism
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / chemistry*
  • Bacterial Toxins / chemistry*
  • Binding Sites
  • Binding, Competitive
  • Cell Membrane / metabolism
  • Dose-Response Relationship, Drug
  • Endotoxins / chemistry*
  • Hemolysin Proteins
  • Immunoblotting
  • Insect Proteins / chemistry*
  • Insecta
  • Kinetics
  • Ligands
  • Microvilli / metabolism
  • Moths
  • Pest Control, Biological
  • Protein Binding
  • Receptors, Cell Surface / chemistry*
  • Surface Plasmon Resonance
  • Time Factors


  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Bacterial Toxins
  • Cry toxin receptors
  • Endotoxins
  • Hemolysin Proteins
  • Insect Proteins
  • Ligands
  • Receptors, Cell Surface
  • insecticidal crystal protein, Bacillus Thuringiensis