Key Residues of Bacillus Thuringiensis Cry2Ab for Oligomerization and Pore-Formation Activity

Abstract As a pore-forming toxin, oligomerization and pore-formation were both required for the mode of action of Cry toxins. Previous studies revealed that Domain I helices α4-α5 were involved in oligomerization of Cry2Ab, while the active residues in charge of Cry2Ab aggregation remained ambiguous. In present studies, we built 20 Cry2Ab alanine mutants site directed in helices α4-α5 and demonstrated that mutants N151A, T152A, F157A, L183A, L185A and I188A blocked the assembly of 250 kDa oligomers, suggesting that those residues were key residues for Cry2Ab oligomerization. As expected, those variants severely reduced the insecticidal activity against P. xylostella which was similar to our previous reports. Furthermore, we found that the pore-forming activities of non-oligomerization mutants sharply decreased compared to wild-type Cry2Ab. Taken together, our data comprehensively identified key residues for Cry2Ab for the first time and emphasized that oligomerization was closely related to insecticidal activity and pore-forming activity in Cry2Ab.