Kinetics of Bacillus thuringiensis toxin binding with brush border membrane vesicles from susceptible and resistant larvae of Plutella xylostella.
An optical biosensor technology based on surface plasmon resonance was used to determine the kinetic rate constants for interactions between the CryIA(c) toxin from Bacillus thuringiensis and brush border membrane vesicles purified from susceptible and resistant larvae of diamondback moth (Plutella xylostella). CryIA(c) association and dissociation rate constants for vesicles from susceptible larvae were determined to be 4.5 x 10(3) M-1 s-1 and 3.2 x 10(-5) s-1, respectively, resulting in a calculated affinity constant of 7 nM. CryIE toxin did not kill susceptible or resistant larvae and did not bind to brush border vesicles. Contrary to expectations based on previous studies of binding in resistant P. xylostella, the binding kinetics for CryIA(c) did not differ significantly between susceptible larvae and those that were resistant to CryIA(c). Determination of the number of CryIA(c) receptors revealed an approximately 3-fold decrease in total CryIA(c) receptor numbers for resistant vesicles. These results suggest that factors other than binding may be altered in our resistant diamondback moth strain. They also support the view that binding is not sufficient for toxicity.
Masson
L
L
Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada.
Mazza
A
A
Brousseau
R
R
Tabashnik
B
B
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D001427:Bacterial Toxins; D015374:Biosensing Techniques; D004731:Endotoxins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D006460:Hemolysin Proteins; D007305:Insecticide Resistance; D007306:Insecticides / Q000378:metabolism*; D007422:Intestines / Q000648:ultrastructure; D007700:Kinetics; D007814:Larva; D008871:Microvilli / Q000378:metabolism*; D009036:Moths / Q000187:drug effects* / Q000254:growth & development; D010572:Pest Control, Biological; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000494:pharmacology
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007306:Insecticides; D011993:Recombinant Fusion Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis