Skip to Main Content
Isolation and partial characterization of gypsy moth BTR-270, an anionic brush border membrane glycoconjugate that binds Bacillus thuringiensis Cry1A toxins with high affinityAuthor(s): Algimantas P. Valaitis; Jeremy L. Jenkins; Mi Kyong Lee; Donald H. Dean; Karen J. Garner
Source: Archives of Insect Biochemistry and Physiology. 46: 186-200.
Publication Series: Scientific Journal (JRNL)
Station: Northern Research Station
PDF: Download Publication (258.94 KB)
DescriptionBTR-270, a gypsy moth (Lymantria dispar) brush border membrane molecule that binds Bacillus thuringiensis (Bt) Cry1A toxins with high affinity, was purified by preparative gel electrophoresis. Rabbit antibodies specific for the Bt toxin-binding molecule were raised. Attempts to label BTR-270 by protein-directed techniques were futile, but it was degraded by proteases with broad specificity indicating the presence of a peptide. Carbohydrate was detected by labeling with digoxigenin hydrazide following periodate oxidation. Mild alkaline hydrolysis destroyed toxin and antibody binding, suggesting O-linked glycans are involved in the activity. GC/MS composition analysis showed that the predominant sugars were galactose, glucose, and N-acetyl galactosamine with lesser amounts of N-acetyl glucosamine, glucuronic acid, xylose, and fucose. The carbohydrate moiety accounted for 73% of its total mass. Amino acid analysis showed a high content of aspartic/asparagine, threonine, and serine residues in the protein moiety. The purified glycoconjugate was not visualized using Coomassie or silver staining procedures, but stained "blue" using the cationic dye Stains-all. BTR-270 was labeled with biotin and used as a diagnostic probe for screening and identifying toxins that bind to the receptor. Toxin-binding kinetics obtained using a biosensor demonstrated that the receptor binds Cry1Aa and Cry1Ab toxins with high affinity, and displays a weaker affinity for Cry1Ac, in correlation with the toxicity of these toxins towards gypsy moth.
- Check the Northern Research Station web site to request a printed copy of this publication.
- Our on-line publications are scanned and captured using Adobe Acrobat.
- During the capture process some typographical errors may occur.
- Please contact Sharon Hobrla, firstname.lastname@example.org if you notice any errors which make this publication unusable.
- We recommend that you also print this page and attach it to the printout of the article, to retain the full citation information.
- This article was written and prepared by U.S. Government employees on official time, and is therefore in the public domain.
CitationValaitis, Algimantas P.; Jenkins, Jeremy L.; Lee, Mi Kyong; Dean, Donald H.; Garner, Karen J. 2001. Isolation and partial characterization of gypsy moth BTR-270, an anionic brush border membrane glycoconjugate that binds Bacillus thuringiensis Cry1A toxins with high affinity. Archives of Insect Biochemistry and Physiology. 46: 186-200.
Keywordsgypsy moth, BTR-270, Bacillus thuringiensis Cry1A toxin receptor
- Localization of Bacillus thuringiensis Cry1A toxin-binding molecules in gypsy moth larval gut sections using fluorescence microscopy
- Histochemical study of lectin binding sites in fourth and fifth instar gypsy moth larval midgut epithelium
- Identification of a non-LTR retrotransposon from the gypsy moth
XML: View XML