Molecular and immunological characterization of wheat Serpin (Tri a 33)

Mol Nutr Food Res. 2012 Dec;56(12):1874-83. doi: 10.1002/mnfr.201200244. Epub 2012 Oct 29.

Abstract

Scope: Several wheat proteins are responsible for food and respiratory allergies. Due to their large polymorphism, the allergenic potential of a number of them has not yet been precisely established. The aim of this work was to perform a thorough assessment of serpin (Tri a 33) allergenicity.

Methods and results: Recombinant wheat Serpin-Z2B isoform (rSerpin-Z2B) was expressed in Escherichia coli. Synchrotron radiation circular dichroism data indicated that the recombinant serpin contains slightly more β-strands than α-helix structures. IgE reactivity of sera from 103 patients with food allergy and 29 patients with Baker's asthma was evaluated using ELISA, a model of basophil activation and linear epitope mapping (Pepscan). Twenty percent of patients with food allergy to wheat and 31% of those with Baker's asthma displayed rSerpin-Z2B-specific IgE in ELISA. The protein was able to induce IgE-dependent basophil degranulation. The Pepscan experiment identified four regions involved in IgE binding to serpin. Heating the protein induced its irreversible denaturation and impaired IgE binding, revealing the predominance of conformational epitopes.

Conclusion: This study confirms wheat serpin allergenicity and shows that recombinant serpin may be a marker of a broad spectrum of sensitization to wheat proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Allergens / adverse effects
  • Allergens / immunology
  • Antigens, Plant / immunology*
  • Case-Control Studies
  • Child
  • Child, Preschool
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Gene Expression
  • Humans
  • Immunoglobulin E / blood
  • Immunoglobulin E / immunology
  • Male
  • Middle Aged
  • Plant Proteins / immunology*
  • Recombinant Proteins / immunology
  • Serpins / immunology*
  • Triticum / chemistry*
  • Triticum / immunology
  • Wheat Hypersensitivity / diagnosis
  • Wheat Hypersensitivity / immunology*
  • Young Adult

Substances

  • Allergens
  • Antigens, Plant
  • Plant Proteins
  • Recombinant Proteins
  • Serpins
  • Immunoglobulin E