Amino acid substitutions in alphaA and alphaC of Cyt2Aa2 alter hemolytic activity and mosquito-larvicidal specificity
| Autor: | Wanwarang Pathaichindachote, Amporn Rungrod, Patcharee Promdonkoy, Chartchai Krittanai, Boonhiang Promdonkoy, Sakol Panyim |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Erythrocytes
Mosquito Control Mutant Bacterial Toxins Molecular Sequence Data Bacillus thuringiensis Bioengineering Aedes aegypti Biology medicine.disease_cause Applied Microbiology and Biotechnology Hemolysis Protein Structure Secondary Hemolysin Proteins Bacterial Proteins Species Specificity medicine Animals Amino Acid Sequence Protein Structure Quaternary chemistry.chemical_classification Sheep Bacillus thuringiensis Toxins Toxin fungi Cell Membrane Wild type General Medicine biology.organism_classification Proteinase K Molecular biology In vitro Amino acid Endotoxins Protein Subunits Culicidae Biochemistry chemistry Amino Acid Substitution Solubility Larva biology.protein Mutagenesis Site-Directed Mutant Proteins Biotechnology |
| Zdroj: | Journal of biotechnology. 133(3) |
| ISSN: | 0168-1656 |
| Popis: | Cyt2Aa2 produced by Bacillus thuringiensis subsp. darmstadiensis exhibits in vitro cytolytic activity against broad range of cells but shows specific in vivo toxicity against larvae of Dipteran insects. To investigate the role of amino acids in alphaA and alphaC of this toxin, 3 single-point mutants (A61C, S108C and V109A) were generated. All 3 mutant proteins were highly produced as inclusion bodies that could be solubilized and activated by proteinase K similar to that of the wild type. Hemolytic activity of A61C and S108C mutants was significantly reduced whereas the V109A mutant showed comparable hemolytic activity to the wild type. Interestingly, the A61C mutant exhibited high larvicidal activity to both Aedes aegypti and Culex quinquefasciatus. S108C and V109A mutants showed low activity against C. quinquefasciatus but relatively high toxicity to A. aegypti. These results demonstrated for the first time that amino acids in alphaA and alphaC are involved in the selectivity of the Cyt toxin to the targeted organism. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect