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http://dx.doi.org/10.5483/BMBRep.2011.44.10.674

Essential role of tryptophan residues in toxicity of binary toxin from Bacillus sphaericus  

Kunthic, Thittaya (Institute of Molecular Biosciences, Mahidol University, Salaya Campus)
Promdonkoy, Boonhiang (National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency)
Srikhirin, Toemsak (Department of Physics, Faculty of Sciences, Mahidol University)
Boonserm, Panadda (Institute of Molecular Biosciences, Mahidol University, Salaya Campus)
Publication Information
BMB Reports / v.44, no.10, 2011 , pp. 674-679 More about this Journal
Abstract
Bacillus sphaericus produces mosquito-larvicidal binary toxin composed of BinA and BinB. While BinB is expected to bind to a specific receptor on the cell membrane, BinA interacts to BinB or BinB receptor complex and translocates into the cytosol to exert its activity via unknown mechanism. To investigate functional roles of aromatic cluster in BinA, amino acids at positions Y213, Y214, Y215, W222 and W226 were substituted by leucine. All mutant proteins were highly produced and their secondary structures were not affected by these substitutions. All mutants are able to insert into lipid monolayers as observed by Langmuir-Blodgett trough and could permeabilize the liposomes in a similar manner as the wild type. However, mosquito-larvicidal activity was abolished for W222L and W226L mutants suggesting that tryptophan residues at both positions play an important role in the toxicity of BinA, possibly involved in the cytopathological process after toxin entry into the cells.
Keywords
Bacillus sphaericus; Binary toxin; Calcein release; Langmuir-blodgett; Membrane insertion;
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