Analysis of Calcium-Induced Conformational Changes in Calcium-Binding Allergens and Quantitative Determination of Their IgE Binding Properties

  • Nuria Parody
  • Miguel Ángel Fuertes
  • Carlos Alonso
  • Yago Pico de Coaña
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 963)

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 Ca2+-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.

Key words

Allergen Polcalcin Calcium EF-hand IgE binding 

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Nuria Parody
    • 1
    • 2
  • Miguel Ángel Fuertes
    • 1
  • Carlos Alonso
    • 1
  • Yago Pico de Coaña
    • 1
  1. 1.Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones CientificasUniversidad Autonoma de MadridMadridSpain
  2. 2.R&D DepartmentLaboratorios LETI S.LMadridSpain

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