Docking-based generation of antibodies mimicking Cry1A/1B protein binding sites as potential insecticidal agents against diamondback moth (Plutella xylostella)
文献类型： 外文期刊
作者： Xie, Yajing 1 ; Xu, Chongxin 1 ; Gao, Meijing 1 ; Zhang, Xiao 1 ; Lu, Lina 1 ; Hu, Xiaodan 1 ; Chen, Wei 1 ; Jurat-Fuente 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Key Lab Control Technol & Stand Agroprod Safety &, Jiangsu Prov State Key Lab Breeding Base,Minist A, Key Lab Food Qual & Safety,Jiangsu Acad Agr Sci, Nanjing, Peoples R China
2.Univ Tennessee, Dept Entomol & Plant Pathol, Knoxville, TN 37901 USA
关键词： Cry insecticidal protein; mimic antibody; docking based generation; insecticidal antibody; Plutella xylostella
期刊名称：PEST MANAGEMENT SCIENCE （ 影响因子：4.845； 五年影响因子：4.674 ）
ISSN： 1526-498X
年卷期： 2021 年 77 卷 10 期
页码：
收录情况： SCI
摘要： BACKGROUND Broad use of insecticidal Cry proteins from Bacillus thuringiensis in biopesticides and transgenic crops has resulted in cases of practical field resistance, highlighting the need for novel approaches to insect control. Previously we described an anti-Cry1Ab idiotypic-antibody (B12-scFv) displaying toxicity against rice leafroller (Cnaphalocrocis medinalis) larvae, supporting the potential of antibodies for pest control. The goal of the present study was to generate insecticidal antibodies against diamondback moth (Plutella xylostella) larvae. RESULTS Four genetically engineered antibodies (GEAbs) were designed in silico from B12-scFv using three-dimensional (3D) structure and docking predictions to alkaline phosphatase (ALP) as a Cry1Ac receptor in P. xylostella. Among these GEAbs, the GEAb-dV(L) antibody consisting of two light chains had overlapping binding sites with Cry1A and Cry1B proteins and displayed high binding affinity to P. xylostella midgut brush border membrane (BBM) proteins. Proteins in BBM identified by pull-down assays as binding to GEAb-dV(L) included an ABC transporter and V-ATPase subunit A protein. Despite lacking the alpha-helical structures in Cry1A that are responsible for pore formation, ingestion of GEAb-dV(L) disrupted the P. xylostella larval midgut epithelium and resulted in toxicity. Apoptotic genes were activated in gut cells upon treatment with GEAb-dV(L). CONCLUSION This study describes the first insecticidal GEAb targeting P. xylostella by mimicking Cry proteins. Data support that GEAb-dV(L) toxicity is associated to activation of intracellular cell death pathways, in contrast to pore-formation associated toxicity of Cry proteins. This work provides a foundation for the design of novel insecticidal antibodies for insect control.
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