Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin

Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry δ-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned...

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Main Authors: Moonsom S., Chaisri U., Kasinrerk W., Angsuthanasombat C.
Format: Article
Language:English
Published: 2014
Online Access:http://www.scopus.com/inward/record.url?eid=2-s2.0-35348834827&partnerID=40&md5=1940b1ec700193614a452d7e4951b41e
http://cmuir.cmu.ac.th/handle/6653943832/710
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-7102014-08-29T09:01:59Z Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin Moonsom S. Chaisri U. Kasinrerk W. Angsuthanasombat C. Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry δ-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a β-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins. 2014-08-29T09:01:59Z 2014-08-29T09:01:59Z 2007 Article 12258687 17927913 http://www.scopus.com/inward/record.url?eid=2-s2.0-35348834827&partnerID=40&md5=1940b1ec700193614a452d7e4951b41e http://cmuir.cmu.ac.th/handle/6653943832/710 English
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
language English
description Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry δ-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a β-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.
format Article
author Moonsom S.
Chaisri U.
Kasinrerk W.
Angsuthanasombat C.
spellingShingle Moonsom S.
Chaisri U.
Kasinrerk W.
Angsuthanasombat C.
Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
author_facet Moonsom S.
Chaisri U.
Kasinrerk W.
Angsuthanasombat C.
author_sort Moonsom S.
title Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
title_short Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
title_full Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
title_fullStr Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
title_full_unstemmed Binding characteristics to mosquito-larval midgut proteins of the cloned domain II-III fragment from the Bacillus thuringiensis Cry4Ba toxin
title_sort binding characteristics to mosquito-larval midgut proteins of the cloned domain ii-iii fragment from the bacillus thuringiensis cry4ba toxin
publishDate 2014
url http://www.scopus.com/inward/record.url?eid=2-s2.0-35348834827&partnerID=40&md5=1940b1ec700193614a452d7e4951b41e
http://cmuir.cmu.ac.th/handle/6653943832/710
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