Watermelon profilin: characterization of a major allergen as a model for plant-derived food profilins

Int Arch Allergy Immunol. 2010;153(3):215-22. doi: 10.1159/000314361. Epub 2010 May 18.

Abstract

Background: Plant profilins have been reported as minor allergens. They are a well-known pan-allergen family responsible for cross-reactivity between plant-derived foods and pollens. Watermelon profilin has been reported to be a major allergen in watermelon (Citrullus lanatus).The aim of this study was to characterize recombinant watermelon profilin, confirming its reactivity for diagnostic purposes and the development of immunotherapy.

Methods: Native profilin was purified from watermelon extract by affinity chromatography using poly-L-proline. Recombinant His-tagged profilin was produced in Pichia pastoris yeast using pPICZαA vector and purified by metal chelate affinity chromatography. ELISA and immunoblot were carried out with sera from 17 watermelon-allergic patients. Biological activity was tested by the basophil activation test.

Results: Native profilin and recombinant profilin were purified and identified by mass spectrometry. Both show similar IgE reactivity in vitro and are biologically active.

Conclusions: Similarities were found in the IgE-binding patterns and biological activity of recombinant profilin and native profilin. Recombinant profilin may be a powerful tool for specific diagnosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Amino Acid Sequence
  • Child
  • Child, Preschool
  • Citrullus / chemistry
  • Citrullus / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Hypersensitivity* / diagnosis
  • Immunoglobulin E / blood
  • Immunoglobulin E / immunology*
  • Male
  • Mass Spectrometry
  • Models, Biological
  • Molecular Sequence Data
  • Pichia / genetics
  • Profilins / genetics
  • Profilins / immunology*
  • Profilins / isolation & purification
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology*
  • Sequence Alignment

Substances

  • Profilins
  • Recombinant Proteins
  • Immunoglobulin E

Associated data

  • GENBANK/AY730591