title: Cytotoxicity and binding profiles of activated Cry1Ac and Cry2Ab to three insect cell lines.
abstract: While Cry1Ac has been known to bind with larval midgut proteins cadherin, APN (amino peptidase N), ALP (alkaline phosphatase) and ABCC2 (adenosine triphosphate-binding cassette transporter subfamily C2), little is known about the receptors of Cry2Ab. To provide a clue to the receptors of Cry2Ab, we tested the baseline cytotoxicity of activated Cry1Ac and Cry2Ab against the midgut and fat body cell lines of Helicoverpa zea and the ovary cell line of Spodoptera frugiperda (SF9). As expected, the descending order of cytotoxicity of Cry1Ac against the three cell lines in terms of 50% lethal concetration (LC50) was midgut (31.0 μg/mL) > fat body (59.0 μg/mL) and SF9 cell (99.6 μg/mL). By contrast, the fat body cell line (LC50= 7.55 μg/mL) was about twice more susceptible to Cry2Ab than the midgut cell line (16.0 μg/mL), the susceptibility of which was not significantly greater than that of SF9 cells (27.0 μg/mL). Further, ligand blot showed the binding differences between Cry1Ac and Cry2Ab in the three cell lines. These results indicated that the receptors of Cry2Ab were enriched in fat body cells and thus largely different from the receptors of Cry1Ac, which were enriched in midgut cells.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.|Department of Entomology and BIO5 Institute, University of Arizona, Tucson, AZ, USA.
authors.forename: Jizhen
authors.initials: J
authors.lastname: Wei
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Gemei
authors.initials: G
authors.lastname: Liang
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Kongming
authors.initials: K
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Gu
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.
authors.forename: Yuyuan
authors.initials: Y
authors.lastname: Guo
authors.affiliation: USDA-ARS, Crop Genetics and Breeding Research Unit, Tifton, GA, USA.
authors.forename: Xinzhi
authors.initials: X
authors.lastname: Ni
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.|Department of Entomology and BIO5 Institute, University of Arizona, Tucson, AZ, USA.
authors.forename: Xianchun
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry; D001426:Bacterial Proteins / Q000378:metabolism; D002460:Cell Line; D004731:Endotoxins / Q000378:metabolism / Q000633:toxicity*; D005216:Fat Body; D041981:Gastrointestinal Tract; D006460:Hemolysin Proteins / Q000378:metabolism / Q000633:toxicity*; D011485:Protein Binding; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins
keywords: Bt; SF9 cell; cell toxicity; fat body cell; midgut cell; receptors