title: Recognition and binding of the PF2 lectin to α-amylase from Zabrotes subfasciatus (Coleoptera:Bruchidae) larval midgut.
abstract: Amylases are an important family of enzymes involved in insect carbohydrate metabolism that are required for the survival of insect larvae. For this reason, enzymes from starch-dependent insects are targets for insecticidal control. PF2 (Olneya tesota) is a lectin that is toxic to Zabrotes subfasciatus (Coleoptera: Bruchidae) larvae. In this study, we evaluated recognition of the PF2 lectin to α-amylases from Z. subfasciatus midgut and the effect of PF2 on α-amylase activity. PF2 caused a decrease of total amylase activity in vitro. Subsequently, several α-amylase isoforms were isolated from insect midgut tissues using ion exchange chromatography. Three enzyme isoforms were verified by an in-gel assay for amylase activity; however, only one isoform was recognized by antiamylase serum and PF2. The identity of this Z. subfasciatus α-amylase was confirmed by liquid chromatography-tandem mass spectrometry. The findings strongly suggest that a glycosylated α-amylase isoform from larval Z. subfasciatus midgut interacts with PF2, which interferes with starch digestion.
authors.affiliation: Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo, A.C. Apartado Postal 1735, Hermosillo, México.
authors.forename: I
authors.initials: I
authors.lastname: Lagarda-Diaz
authors.affiliation: Department of Nutritional Sciences, University of Arizona, Tucson, AZ 85721.
authors.forename: D
authors.initials: D
authors.lastname: Geiser
authors.affiliation: Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo, A.C. Apartado Postal 1735, Hermosillo, México.
authors.forename: A M
authors.initials: AM
authors.lastname: Guzman-Partida
authors.affiliation: Department of Nutritional Sciences, University of Arizona, Tucson, AZ 85721.
authors.forename: J
authors.initials: J
authors.lastname: Winzerling
authors.affiliation: Coordinación de Ciencia de los Alimentos, Centro de Investigación en Alimentación y Desarrollo, A.C. Apartado Postal 1735, Hermosillo, México lvazquez@ciad.mx.
authors.forename: L
authors.initials: L
authors.lastname: Vazquez-Moreno
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000502:physiology*; D004064:Digestive System / Q000187:drug effects* / Q000378:metabolism; D007887:Fabaceae / Q000737:chemistry* / Q000378:metabolism; D007814:Larva / Q000187:drug effects / Q000502:physiology; D037102:Lectins / Q000378:metabolism*; D037121:Plant Lectins; D010940:Plant Proteins / Q000378:metabolism*; D000516:alpha-Amylases / Q000037:antagonists & inhibitors / Q000096:biosynthesis*
chemical_list: D037102:Lectins; D037121:Plant Lectins; D010940:Plant Proteins; D000516:alpha-Amylases; C057084:glycosylated alpha-amylase
keywords: PF2; Zabrotes subfasciatus; amylase; insecticidal effect; lectin