title: Analysis of calcium-induced conformational changes in calcium-binding allergens and quantitative determination of their IgE binding properties.
abstract: The polcalcin family is one of the most epidemiologically relevant families of calcium-binding allergens. Polcalcins are potent plant allergens that contain one or several EF-hand motifs and their allergenicity is primarily associated with the Ca(2+)-bound form of the protein. Conformation, stability, as well as IgE recognition of calcium-binding allergens greatly depend on the presence of protein-bound calcium ions. We describe a protocol that uses three techniques (SDS-PAGE, circular dichroism spectroscopy, and ELISA) to describe the effects that calcium has on the structural changes in an allergen and its IgE binding properties.
authors.affiliation: Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas, Universidad Autonoma de Madrid, Madrid, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Parody
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Fuertes
authors.forename: Carlos
authors.initials: C
authors.lastname: Alonso
authors.forename: Yago
authors.initials: Y
authors.lastname: Pico de Coaña
mesh_terms: D052179:Antigens, Plant / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D002118:Calcium / Q000494:pharmacology*; D029781:Cupressus; D004591:Electrophoresis, Polyacrylamide Gel; D007073:Immunoglobulin E / Q000276:immunology*; D011487:Protein Conformation / Q000187:drug effects
chemical_list: D052179:Antigens, Plant; D007073:Immunoglobulin E; D002118:Calcium