Applied and Environmental Microbiology (2017) 83 (4 - e02864-16)

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Zishan Zhou, Yuxiao Liu, Gemei Liang, Yongping Huang, Alejandra Bravo, Mario Soberón, Fuping Song, Xueping Zhou and Jie Zhang (2017)
Insecticidal specificity of Cry1Ah to Helicoverpa armigera is determined by binding of APN1 via domain II loops 2 and 3
Applied and Environmental Microbiology 83 (4 - e02864-16)
Abstract: Bacillus thuringiensis Cry1Ah protein is highly toxic against Helicoverpa armigera but shows no toxicity against Bombyx mori larvae. In contrast, the closely related Cry1Ai toxin showed the opposite phenotype: high activity against B. mori but no toxicity against H. armigera. Analysis of binding of Cry1Ah to brush border membrane vesicle (BBMV) proteins from H. armigera and B. mori by surface plasmon resonance revealed association of toxin binding with insect specificity. Pulldown experiments identified aminopeptidase N1 (APN1) as a Cry1Ah binding protein that was not observed in the assays using B. mori BBMV proteins. The APN1 Cry1Ah binding region was narrowed to the region from ^A548 to ^S798 (fragment H3) by expressing four different APN1 fragments in Escherichia coli and analyzing Cry1Ah binding by ligand blot. Binding competition experiments of Cry1Ah to APN1 fragment H3 using synthetic peptides corresponding to four predicted domain II loop regions showed that loop 2 and loop 3 have additive effects on binding to APN1 fragment H3. Moreover, switching of loop 2 and loop 3 regions from Cry1Ah to Cry1Ai toxins showed that loop 2 and loop 3 are both involved in specificity and toxicity against H. armigera.
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Database assignments for author(s): Jie Zhang, Alejandra Bravo, Ge-Mei Liang, Yongping Huang

Research topic(s) for pests/diseases/weeds:
biocontrol - natural enemies
Research topic(s) for beneficials or antagonists:
molecular biology - genes
general biology - morphology - evolution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Helicoverpa armigera
Bacillus thuringiensis Cry1A-toxin (entomopathogen)	Helicoverpa armigera