Ligand blot identification of a Manduca sexta midgut binding protein specific to three Bacillus thuringiensis CryIA-type ICPs.
The CryIA(a), CryIA(b) and CryIA(c) Bacillus thuringiensis insecticidal crystal proteins (ICPs) were used in ligand-blot experiments to detect specific binding proteins in brush-border membrane vesicles (BBMV) of Manduca sexta. We identified a protein which binds these three CryIA-type ICPs. The apparent molecular mass of the protein, estimated on SDS-PAGE, was 210 kDa as was the CryIA(b) binding protein previously described by Vadlamudi and col. We have also demonstrated, in ligand blot experiments, that CryIA(a) and CryIA(c) compete with CryIA(b) for binding this 210 kDa protein. Properties of the binding molecule can be correlated with knowledge previously acquired through radiolabelled binding experiments.
Martínez-Ramírez
A C
AC
Department of Genetics, University of Valencia, Spain.
González-Nebauer
S
S
Escriche
B
B
Real
M D
MD
D000818:Animals; D001413:Bacillus thuringiensis; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D001667:Binding, Competitive; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D004064:Digestive System / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000378:metabolism*; D006460:Hemolysin Proteins; D007457:Iodine Radioisotopes; D008871:Microvilli / Q000378:metabolism; D008970:Molecular Weight; D009036:Moths / Q000378:metabolism*; D011869:Radioligand Assay
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D002352:Carrier Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; D007457:Iodine Radioisotopes; C030956:insecticidal crystal protein, Bacillus Thuringiensis