Production and characterization of N- and C-terminally truncated Mtx2: a mosquitocidal toxin from Bacillus sphaericus.
Mosquitocidal toxin 2 (Mtx2) is a mosquito-larvicidal protein produced during vegetative stage of Bacillus sphaericus. The toxin consists of 292 amino acids and has a molecular weight of 31.8 kDa. To determine the active core region of the toxin, amino acids at N- and C-termini were sequentially removed. Deletion up to 23 amino acids from the N-terminus (Met1-Tyr23) did not significantly affect protein production and the toxin activity, whereas removal of 26 amino acids from the N-terminus (Met1-Lys26) completely abolished toxicity even though the protein production remained unchanged. Deletion of only 5 amino acids from the C-terminal end yielded the protein that could not be solubilized and rendered the toxin inactive. The results demonstrated that the C-terminal end of Mtx2 is required for proper folding and toxicity. Amino acids at the N-terminus up to Tyr23 did not play a significant role in protein production and toxicity whereas amino acids between Thr24 and Lys26 are required for full toxicity.
Phannachet
Kulwadee
K
Department of Biotechnology, Faculty of Science and Technology, Thammasat University, 99 Pahonyothin Road, Klong 1, Klong Luang, Pathumthani, 12120, Thailand.
Raksat
Ponlawoot
P
Limvuttegrijeerat
Thidarat
T
Promdonkoy
Boonhiang
B
D000818:Animals; D001407:Bacillus / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*; D009033:Culicidae / Q000187:drug effects; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D017510:Protein Folding; D017384:Sequence Deletion
D001426:Bacterial Proteins; D001427:Bacterial Toxins; C099210:Mtx2 protein, Bacillus sphaericus; D050505:Mutant Proteins
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