Aminopeptidase N from Bombyx Mori as a Candidate for the Receptor of Bacillus Thuringiensis Cry1Aa Toxin
Open Access
- 1 June 1997
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 246 (3) , 652-657
- https://doi.org/10.1111/j.1432-1033.1997.t01-1-00652.x
Abstract
Cry1Aa toxin‐binding proteins from the midgut brush border membrane vesicles of Bombyx mori, a toxin‐susceptible silkworm, were analyzed to find candidates for the toxin receptors. Ligand blotting showed that Cry1Aa toxin bound to a 120‐kDa protein. A part of the 120‐kDa protein was solubilized from the membrane vesicles with phosphatidylinositol‐specific phospholipase C, resulting in a 110‐kDa protein which therefore may be linked to a glycosylphosphatidylinositol anchor. The 120‐kDa and 110‐kDa Cry1Aa toxin‐binding proteins were solubilized with detergent or pohosphatidylinositol‐specific phospholipase C, respectively, and purified using anion‐exchange chromatography. Scatchard plot analysis for the specific binding of purified 110‐kDa protein to Cry1Aa toxin yielded a Kd value of 7.6 nM, which was similar to that for the binding of intact brush border membrane vesicles to the toxin. N‐terminal and internal amino acid sequences of the 120‐kDa and 110‐kDa proteins showed high degrees of similarity to those of aminopeptidase N, a putative Cry1Ac toxin receptor, reported in Manduca sexta and Heliothis virescens. On this basis, the 120‐kDa Cry1Aa toxin‐binding protein from B. mori was identified as a member of the aminopeptidase family.Keywords
This publication has 49 references indexed in Scilit:
- Identification, Isolation, and Cloning of a Bacillus thuringiensis CryIAc Toxin-binding Protein from the Midgut of the Lepidopteran Insect Heliothis virescensPublished by Elsevier ,1995
- δ‐Endotoxins induce cation channels in Spodoptera frugiperda brush border membranes in suspension and in planar lipid bilayersFEBS Letters, 1995
- The receptor for Bacillus thuringiensis CrylA(c) delta‐endotoxin in the brush border membrane of the lepidopteran Manduca sexta is aminopeptidase NMolecular Microbiology, 1994
- The toxicity of twoBacillus thuringiensis δ-endotoxins to gypsy moth larvae is inversely related to the affinity of binding sites on midgut brush border membranes for the toxinsCellular and Molecular Life Sciences, 1990
- A cytolytic δ‐endotoxin from Bacillus thuringiensis var. israelensis forms cation‐selective channels in planar lipid bilayersFEBS Letters, 1989
- Evaluation of equilibrium constants for antigen-antibody interactions by solid-phase immunoassay: The binding of paraquat to its elicited mouse monoclonal antibodyMolecular Immunology, 1987
- A toxic fragment from the entomocidal crystal protein of Bacillus thuringiensis.Agricultural and Biological Chemistry, 1984
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- THE ATTRACTIONS OF PROTEINS FOR SMALL MOLECULES AND IONSAnnals of the New York Academy of Sciences, 1949