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Bacillus thuringiensis toxin, Cry1C interacts with 128HLHFHLP134 region of aminopeptidase N of agricultural pest, Spodoptera litura

Paper ID Volume ID Publish Year Pages File Format Full-Text
34717 45040 2014 9 PDF Available
Title
Bacillus thuringiensis toxin, Cry1C interacts with 128HLHFHLP134 region of aminopeptidase N of agricultural pest, Spodoptera litura
Abstract

•In silico docking and biopanning identified a putative binding region of Cry1C 128HLHFHLP134 to APN of Spodoptera litura.•Synthetic peptides homologous to domain II loop 2 and 3 of Cry1C competed Cry1C-APN binding.•Alanines substitution of H128, H130, H132 and P134 affect the toxicity of Cry1C toxin to APN receptor of S. litura.

We modeled Cry1C toxin and its Aminopeptidase-N receptor and in silico docking analysis was performed. Further, we utilized biopanning against Cry1C followed by blocking assays and mutagenesis analysis to identify the binding epitope of SlAPN. We have identified a putative SlAPN binding region, APN-CRY (128HLHFHLP134). A derivative of SlAPN carrying the 128HLHFHLP134 region termed as binding region of APN (BR-APN) was cloned and its involvement in Cry1C binding and toxicity was checked. Cry1C-BR-APN binding was competed by synthetic peptides homologous to loop2 and loop3 of domain II but not by that of loopα. Additionally, alanines substitution of residues H128, H130, H132 and P134 affect the binding efficiency of receptor to Cry1C toxin (upto 4-fold lower affinity).These residues are also implicated in Cry1C toxicity as shown by the reduced ability to affect the mortality of Cry1C on S. litura larvae when toxin was preincubated with a fragment of the receptor.

Keywords
Agriculture pest; Spodoptera litura; Cry1C; Aminopeptidase1; Toxin–receptor interaction; MutagenesisBt, Bacillus thuringiensis; SlAPN, recombinantly expressed APN of Spodoptera litura; BBMV, brush border membrane vesicle
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Bacillus thuringiensis toxin, Cry1C interacts with 128HLHFHLP134 region of aminopeptidase N of agricultural pest, Spodoptera litura
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 49, Issue 4, April 2014, Pages 688–696
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us