Journal
BMB REPORTS
Volume 44, Issue 10, Pages 674-679Publisher
KOREAN SOCIETY BIOCHEMISTRY & MOLECULAR BIOLOGY
DOI: 10.5483/BMBRep.2011.44.10.674
Keywords
Bacillus sphaericus; Binary toxin; Calcein release; Langmuir-blodgett; Membrane insertion
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Funding
- National Center for Genetic Engineering and Biotechnology
- National Science and Technology Development Agency, Thailand
- Thailand Research Fund
- Mahidol University, Thailand
- Ministry of Science and Technology, Thailand
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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 pemteabilize 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. [BMB reports 2011; 44(10): 674-679]
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