Proteolytic processing of the mosquitocidal toxin from Bacillus sphaericus SSII-1.
The 97-kDa protein Mtx21, derived from the 100-kDa mosquitocidal protein (Mtx) from Bacillus sphaericus SSII-1 by the deletion of the putative signal sequence, was expressed as a fusion protein with glutathione S-transferase in Escherichia coli, and the fusion protein was purified by affinity chromatography. The fusion protein bound to glutathione agarose was cleaved with thrombin to release the Mtx21 protein. The 97-kDa Mtx21 protein was found to be toxic to Culex quinquefasciatus larvae with a 50% lethal concentration of 15 ng/ml. Treating Mtx21 with crude mosquito larval gut extracts gave rise to two major peptides of 70 and 27 kDa. Treating the 97-kDa Mtx21 protein with trysin also gave rise to a similar proteolytic cleavage pattern. N-terminal sequencing showed that the 27-kDa peptide was derived from the N-terminal region of the 97-kDa protein and that the 70-kDa protein was from the C-terminal region of the 97-kDa protein. The 27-kDa peptide has all the previously identified regions of homology with the catalytic peptides of the ADP-ribosyltransferase toxins, such as pertussis toxin S1 peptide, while the 70-kDa peptide has three internal regions of homology.
Insecticidal Toxins Laboratory, National University of Singapore.
T
T
Thanabalu
J
J
Hindley
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C
Berry
D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000737:chemistry; D001427:Bacterial Toxins / Q000032:analysis / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D009033:Culicidae / Q000187:drug effects*; D007814:Larva / Q000187:drug effects; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000032:analysis; D021382:Protein Sorting Signals / Q000032:analysis; D014357:Trypsin / Q000494:pharmacology
D001427:Bacterial Toxins; D010446:Peptide Fragments; D021382:Protein Sorting Signals; C068603:mosquitocidal toxin SSII-1; D011065:Poly(ADP-ribose) Polymerases; D014357:Trypsin