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http://dx.doi.org/10.5483/BMBRep.2014.47.10.192

Functional characterizations of residues Arg-158 and Tyr-170 of the mosquito-larvicidal Bacillus thuringiensis Cry4Ba  

Leetachewa, Somphob (Bacterial Protein Toxin Research Cluster, Institute of Molecular Biosciences, Mahidol University)
Moonsom, Saengduen (Department of Protozoology, Faculty of Tropical Medicine, Mahidol University)
Chaisri, Urai (Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University)
Khomkhum, Narumol (Department of Protozoology, Faculty of Tropical Medicine, Mahidol University)
Yoonim, Nonglak (Faculty of Medical Technology, Western University)
Wang, Ping (Department of Entomology, New York State Agricultural Experiment Station, Cornell University)
Angsuthanasombat, Chanan (Bacterial Protein Toxin Research Cluster, Institute of Molecular Biosciences, Mahidol University)
Publication Information
BMB Reports / v.47, no.10, 2014 , pp. 546-551 More about this Journal
Abstract
The insecticidal activity of Bacillus thuringiensis (Bt) Cry toxins involves toxin stabilization, oligomerization, passage across the peritrophic membrane (PM), binding to midgut receptors and pore-formation. The residues Arg-158 and Tyr-170 have been shown to be crucial for the toxicity of Bt Cry4Ba. We characterized the biological function of these residues. In mosquito larvae, the mutants R158A/E/Q (R158) could hardly penetrate the PM due to a significantly reduced ability to alter PM permeability; the mutant Y170A, however, could pass through the PM, but degraded in the space between the PM and the midgut epithelium. Further characterization by oligomerization demonstrated that Arg-158 mutants failed to form correctly sized high-molecular weight oligomers. This is the first report that Arg-158 plays a role in the formation of Cry4Ba oligomers, which are essential for toxin passage across the PM. Tyr-170, meanwhile, is involved in toxin stabilization in the toxic mechanism of Cry4Ba in mosquito larvae.
Keywords
Aedes aegypti; Bacillus thuringiensis; Cry4Ba toxin; Larvicidal activity; Peritrophic membrane;
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