Role of cysteine at positions 67, 161 and 241 of a Bacillus sphaericus binary toxin BinB.
Binary toxin consisting of BinA and BinB from Bacillus sphaericus is toxic to mosquito larvae. BinB is responsible for specific binding to the larval gut cell membrane while BinA is crucial for toxicity. To investigate functional role of cysteine in BinB, three cysteine residues at positions 67, 161, and 241 were replaced by alanine or serine. Mutations at these positions did not affect protein production and overall structure of BinB. These cysteine residues are not involved in disulfide bond formation between BinB molecules. Mosquito-larvicidal assays revealed that C67 and C161 are essential for toxicity, whereas C241 is not. Mutations at C67 and C161 resulted in weaker BinA-BinB interaction. The loss of toxicity may be due to the reduction of interactions between BinA and BinB or BinB and its receptor. C67 and C161 could also play a part during conformational changes or internalization of the binary toxin into the target cell. [BMB reports 2010; 43(1): 23-28].
Institute of Molecular Biosciences, Mahidol University, Salaya Campus, Putthamonthon 4 Road, Nakhonpathom 73170, Thailand.
Patcharaporn
P
Boonyos
Sumarin
S
Soonsanga
Panadda
P
Boonserm
Boonhiang
B
Promdonkoy
D019943:Amino Acid Substitution; D000818:Animals; D001407:Bacillus / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D009033:Culicidae / Q000187:drug effects; D003545:Cysteine / Q000737:chemistry*; D007814:Larva / Q000187:drug effects; D009032:Mosquito Control; D016297:Mutagenesis, Site-Directed
D001427:Bacterial Toxins; C046134:binB protein, Bacillus sphaericus; D003545:Cysteine