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Review
. 2008 Feb;29(2):324-9.
doi: 10.1016/j.peptides.2007.07.035. Epub 2007 Dec 14.

Employing phage display to study the mode of action of Bacillus thuringiensis Cry toxins

Luisa Elena Fernández  1 , Isabel Gómez, Sabino Pacheco, Iván Arenas, Sarjeet S Gilla, Alejandra Bravo, Mario Soberón
Affiliations
Review

Employing phage display to study the mode of action of Bacillus thuringiensis Cry toxins

Luisa Elena Fernández et al. Peptides. 2008 Feb.

Abstract

Phage display is an in vitro method for selecting polypeptides with desired properties from a large collection of variants. The insecticidal Cry toxins produced by Bacillus thuringiensis are highly specific to different insects. Various proteins such as cadherin, aminopeptidase-N (APN) and alkaline phosphatase (ALP) have been characterized as potential Cry-receptors. We used phage display to characterize the Cry toxin-receptor interaction(s). By employing phage-libraries that display single-chain antibodies (scFv) from humans or from immunized rabbits with Cry1Ab toxin or random 12-residues peptides, we have identified the epitopes that mediate binding of lepidopteran Cry1Ab toxin with cadherin and APN receptors from Manduca sexta and the interaction of dipteran Cry11Aa toxin with the ALP receptor from Aedes aegypti. Finally we displayed in phages the Cry1Ac toxin and discuss the potential for selecting Cry variants with improved toxicity or different specificity.

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Figures

Figure 1
Figure 1
Cry1Ab toxin binds Manduca sexta cadherin repeat 7 by means of domain II loop 2.
Figure 2
Figure 2
Binding of Cry1Ab toxin to the Manduca sexta cadherin receptor promotes cleavage of helix α-1 in domain I and the subsequent formation of a pre-pore oligomeric structure that is membrane-insertion-competent.
Figure 3
Figure 3
The Cry1Ab pre-pore oligomer binds aminopeptidase-N (APN) from Manduca sexta by means of β-16 in domain III.
Figure 4
Figure 4
The Cry11Aa toxin binds Aedes aegypti alkaline phosphatase (ALP) by means of the α-8 loop in domain II.
See this image and copyright information in PMC

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