Intramolecular proteolytic nicking and binding of Bacillus thuringiensis Cry8Da toxin in BBMVs of Japanese beetle

Journal of Invertebrate Pathology
Takuya YamaguchiShin-ichiro Asano

Abstract

Bacillus thuringiensis (Bt) Cry8D insecticidal proteins are unique among Cry8 family proteins in terms of its insecticidal activity against adult Scarab beetles, such as Japanese beetle (Popillia japonica Newman). From the sequence homology with other Bt Cry proteins especially those active against beetles, such as Cry3Aa whose 3D structure is available, the structure of the Cry8D protein has been predicted to be a typical three-domain Cry protein type. In addition, the activation process of Cry8D in gut juice of susceptible insects is presumed to be similar to that of Cry3A (Yamaguchi et al., 2008). In this study, the activation process of Cry8Da in insect gut juice was closely examined. Japanese beetle gut juice proteases digested the 130kDa Cry8Da protein to produce a 64kDa protein. This 64kDa protein was active against both adult and larval Japanese beetle and considered to be an activated toxin. N-terminal sequencing of this 64kDa protein revealed that the Cry8Da leader sequence consisting of 63 amino acid residues from M(1) to F(63) was removed. As in the case of Cry3Aa, the proteases further digested the 64kDa protein to two 8kDa and 54kDa fragments. N-terminal amino acid analysis of these smaller fragments indicated tha...Continue Reading

References

Apr 5, 2007·Nature Protocols·Hermann Schägger
Jan 9, 2009·Current Microbiology·Changlong ShuFuping Song
Apr 29, 2009·Applied Microbiology and Biotechnology·Changlong ShuFuping Song
Oct 13, 2009·Applied and Environmental Microbiology·Carlos Muñoz-GarayAlejandra Bravo

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