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Ligand specificity and affinity of BT-R1, the Bacillus thuringiensis Cry1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures.
Appl Environ Microbiol. 1997 Sep; 63(9):3419-25.AE

Abstract

The Manduca sexta receptor for the Bacillus thuringiensis Cry1Aa, Cry1Ab, and Cry1Ac toxins, BT-R1, has been expressed in heterologous cell culture, and its ligand binding characteristics have been determined. When transfected with the BT-R1 cDNA, insect and mammalian cell cultures produce a binding protein of approximately 195 kDa, in contrast to natural BT-R1 from M. sexia, which has an apparent molecular weight of 210 kDa. Transfection of cultured Spodoptera frugiperda cells with the BT-R1 cDNA imparts Cry1A-specific high-affinity binding activity typical of membranes prepared from larval M. sexta midguts. Competition assays with BT-R1 prepared from larval M. sexta midguts and transiently expressed in cell culture reveal virtually identical affinities for the Cry1Aa, Cry1Ab, and Cry1Ac toxins, clearly demonstrating the absolute specificity of the receptor for toxins of the lepidopteran-specific Cry1A family. BT-R1 therefore remains the only M. sexta Cry1A binding protein to be purified, cloned, and functionally expressed in heterologous cell culture, and for the first time, we are able to correlate the Cry1Aa, Cry1Ab, and Cry1Ac toxin sensitivities of M. sexta to the identity and ligand binding characteristics of a single midgut receptor molecule.

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Authors+Show Affiliations

Keeton TP
Department of Molecular Biology, University of Wyoming, Laramie 82071, USA.
Bulla LA
No affiliation info available

MeSH

AnimalsBacillus thuringiensisBacillus thuringiensis ToxinsBacterial ProteinsBacterial ToxinsBinding, CompetitiveCOS CellsCell LineEndotoxinsGene ExpressionHemolysin ProteinsHumansInsect ProteinsLigandsManducaReceptors, Cell SurfaceRecombinant ProteinsSpodopteraTransfection

Pub Type(s)

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

Language

eng

PubMed ID

9292994

Citation

Keeton, T P., and L A. Bulla. "Ligand Specificity and Affinity of BT-R1, the Bacillus Thuringiensis Cry1A Toxin Receptor From Manduca Sexta, Expressed in Mammalian and Insect Cell Cultures." Applied and Environmental Microbiology, vol. 63, no. 9, 1997, pp. 3419-25.
Keeton TP, Bulla LA. Ligand specificity and affinity of BT-R1, the Bacillus thuringiensis Cry1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures. Appl Environ Microbiol. 1997;63(9):3419-25.
Keeton, T. P., & Bulla, L. A. (1997). Ligand specificity and affinity of BT-R1, the Bacillus thuringiensis Cry1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures. Applied and Environmental Microbiology, 63(9), 3419-25.
Keeton TP, Bulla LA. Ligand Specificity and Affinity of BT-R1, the Bacillus Thuringiensis Cry1A Toxin Receptor From Manduca Sexta, Expressed in Mammalian and Insect Cell Cultures. Appl Environ Microbiol. 1997;63(9):3419-25. PubMed PMID: 9292994.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Ligand specificity and affinity of BT-R1, the Bacillus thuringiensis Cry1A toxin receptor from Manduca sexta, expressed in mammalian and insect cell cultures. AU - Keeton,T P, AU - Bulla,L A,Jr PY - 1997/9/18/pubmed PY - 2001/3/28/medline PY - 1997/9/18/entrez SP - 3419 EP - 25 JF - Applied and environmental microbiology JO - Appl Environ Microbiol VL - 63 IS - 9 N2 - The Manduca sexta receptor for the Bacillus thuringiensis Cry1Aa, Cry1Ab, and Cry1Ac toxins, BT-R1, has been expressed in heterologous cell culture, and its ligand binding characteristics have been determined. When transfected with the BT-R1 cDNA, insect and mammalian cell cultures produce a binding protein of approximately 195 kDa, in contrast to natural BT-R1 from M. sexia, which has an apparent molecular weight of 210 kDa. Transfection of cultured Spodoptera frugiperda cells with the BT-R1 cDNA imparts Cry1A-specific high-affinity binding activity typical of membranes prepared from larval M. sexta midguts. Competition assays with BT-R1 prepared from larval M. sexta midguts and transiently expressed in cell culture reveal virtually identical affinities for the Cry1Aa, Cry1Ab, and Cry1Ac toxins, clearly demonstrating the absolute specificity of the receptor for toxins of the lepidopteran-specific Cry1A family. BT-R1 therefore remains the only M. sexta Cry1A binding protein to be purified, cloned, and functionally expressed in heterologous cell culture, and for the first time, we are able to correlate the Cry1Aa, Cry1Ab, and Cry1Ac toxin sensitivities of M. sexta to the identity and ligand binding characteristics of a single midgut receptor molecule. SN - 0099-2240 UR - https://www.unboundmedicine.com/medline/citation/9292994/Ligand_specificity_and_affinity_of_BT_R1_the_Bacillus_thuringiensis_Cry1A_toxin_receptor_from_Manduca_sexta_expressed_in_mammalian_and_insect_cell_cultures_ L2 - https://journals.asm.org/doi/10.1128/aem.63.9.3419-3425.1997?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -
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