Employing phage display to study the mode of action of Bacillus thuringiensis Cry toxins
Employing phage display to study the mode of action of Bacillus thuringiensis Cry toxins
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DOI：
10.1016/j.peptides.2007.07.035
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发表时间：
2008-02-01
期刊：
影响因子：
3
通讯作者：
Soberon, Mario
中科院分区：
文献类型：
--
作者：
Elena Fernandez, Luisa;Gomez, Isabel;Soberon, Mario
噬菌体展示是用于从大量变体集合中选择具有所需性质的多肽的体外方法。由苏云金芽孢杆菌产生的杀虫Cry毒素对不同的昆虫具有高度特异性。各种蛋白质如钙粘蛋白、氨肽酶-N（APN）和碱性磷酸酶（ALP）已被表征为潜在的Cry受体。我们使用噬菌体展示来表征Cry毒素-受体相互作用。通过采用噬菌体库，展示单链抗体（scFv）从人或免疫兔与Cry 1Ab毒素或随机12-残基肽，我们已经确定了介导的表位，鳞翅目Cry 1Ab毒素与钙粘蛋白和APN受体从天蛾和双翅目Cry 11 Aa毒素与埃及伊蚊ALP受体的相互作用。最后，我们展示了Cry 1Ac毒素，并讨论了选择具有改善的毒性或不同特异性的Cry变体的潜力。(c)2007年爱思唯尔公司All rights reserved.
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. (c) 2007 Elsevier Inc. All rights reserved.