title: Identification of Aminopeptidase-N2 as a Cry2Ab binding protein in Manduca sexta.
abstract: Bacillus thuringiensis Cry2Ab toxin has been used in combination with Cry1Ac for resistance management on the Bt-cotton that is widely planted worldwide. However, little is known regarding Cry2Ab mode of action. Particularly, there is a gap of knowledge on the identification of insect midgut proteins that bind Cry2Ab and mediate toxicity. In the case of Cry1Ab toxin, a transmembrane cadherin protein and glycosyl-phosphatidylinositol (GPI) anchored proteins like aminopeptidase-N1 (APN1) or alkaline-phosphatase (ALP) from Manduca sexta, have been shown to be important for oligomer formation and insertion into the membrane. Binding competition experiments showed that Cry2Ab toxin does not share binding sites with Cry1Ab toxin in M. sexta brush border membrane vesicles (BBMV). Also, that Cry2Ab shows reduced binding to the Cry1Ab binding molecules cadherin, APN1 or ALP. Finally, ligand blot experiments and protein sequence by LC-MS/MS identified APN2 isoform as a Cry2Ab binding protein. Cloning and expression of APN2 confirmed that APN2 is a Cry2Ab binding protein.
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Janette
authors.initials: J
authors.lastname: Onofre
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Meztlli O
authors.initials: MO
authors.lastname: Gaytán
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Arlen
authors.initials: A
authors.lastname: Peña-Cardeña
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Blanca I
authors.initials: BI
authors.lastname: García-Gomez
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Sabino
authors.initials: S
authors.lastname: Pacheco
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Isabel
authors.initials: I
authors.lastname: Gómez
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Bravo
authors.affiliation: Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca 62250, Morelos, Mexico. Electronic address: mario@ibt.unam.mx.
authors.forename: Mario
authors.initials: M
authors.lastname: Soberón
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D018826:CD13 Antigens / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004731:Endotoxins / Q000737:chemistry / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000737:chemistry / Q000378:metabolism*; D019476:Insect Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007305:Insecticide Resistance; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008024:Ligands; D018402:Manduca / Q000201:enzymology* / Q000235:genetics; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; C095416:Cry toxin receptors; D004731:Endotoxins; D006460:Hemolysin Proteins; D019476:Insect Proteins; D007527:Isoenzymes; D008024:Ligands; D011956:Receptors, Cell Surface; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D018826:CD13 Antigens
keywords: Bacillus thuringiensis; Cry toxins; Cry-receptors; GPI-anchored proteins; Gene stacking; Transgenic crops