High-level expression in Escherichia coli and purification of recombinant plant profilins: comparison of IgE-binding capacity and allergenic activity

Biochem Biophys Res Commun. 1996 Sep 4;226(1):42-50. doi: 10.1006/bbrc.1996.1309.

Abstract

Because of their structural similarity and ubiquitous distribution as actin binding proteins, plant profilins represent important cross-reactive allergens for almost 20% of patients suffering from Type I allergy to pollen and other plant products. The cDNAs coding for three birch profilin variants (Tyr44, Glu47, and Asn47), timothy grass profilin, and three tobacco profilin isoforms (ntprof1-3) were expressed at high levels in Escherichia coli as non-fusion proteins. The recombinant plant profilins were purified to homogeneity by poly (L-proline) affinity chromatography and showed comparable capacity to bind IgE-antibodies from profilin allergic patients. All recombinant plant profilins elicited dose-dependent histamine release from basophils of a profilin allergic patient and induced immediate type skin reactions. It is concluded that profilins from different plant species share IgE-epitopes and allergenic properties. Plant profilins therefore constitute a family of functional pan-allergens which may substitute each other for diagnosis and specific immunotherapy.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / genetics*
  • Allergens / isolation & purification
  • Allergens / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites, Antibody
  • Cloning, Molecular
  • Contractile Proteins*
  • Cross Reactions
  • DNA, Complementary
  • Escherichia coli / genetics
  • Histamine Release
  • Humans
  • Immunoglobulin E / metabolism
  • Microfilament Proteins / genetics*
  • Microfilament Proteins / isolation & purification
  • Microfilament Proteins / metabolism
  • Molecular Sequence Data
  • Plant Proteins / genetics*
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism
  • Profilins
  • Protein Binding
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Skin Tests
  • Species Specificity

Substances

  • Allergens
  • Contractile Proteins
  • DNA, Complementary
  • Microfilament Proteins
  • Plant Proteins
  • Profilins
  • Recombinant Proteins
  • Immunoglobulin E