Molecular characterization of recombinant T1, a non-allergenic periwinkle (Catharanthus roseus) protein, with sequence similarity to the Bet v 1 plant allergen family

Biochem J. 2003 Jul 1;373(Pt 1):261-9. doi: 10.1042/BJ20030331.

Abstract

More than 25% of the population suffer from Type I allergy, an IgE-mediated hypersensitivity disease. Allergens with homology to the major birch ( Betula verrucosa ) pollen allergen, Bet v 1, belong to the most potent elicitors of IgE-mediated allergies. T1, a cytokinin-inducible cytoplasmic periwinkle ( Catharanthus roseus ) protein, with significant sequence similarity to members of the Bet v 1 plant allergen family, was expressed in Escherichia coli. Recombinant T1 (rT1) did not react with IgE antibodies from allergic patients, and failed to induce basophil histamine release and immediate-type skin reactions in Bet v 1-allergic patients. Antibodies raised against purified rT1 could be used for in situ localization of natural T1 by immunogold electron microscopy, but did not cross-react with most of the Bet v 1-related allergens. CD analysis showed significant differences regarding secondary structure and thermal denaturation behaviour between rT1 and recombinant Bet v 1, suggesting that these structural differences are responsible for the different allergenicity of the proteins. T1 represents a non-allergenic member of the Bet v 1 family that may be used to study structural requirements of allergenicity and to engineer hypo-allergenic plants by replacing Bet v 1-related allergens for primary prevention of allergy.

Publication types

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

MeSH terms

  • Allergens / chemistry*
  • Allergens / genetics
  • Amino Acid Sequence
  • Antigens, Plant
  • Base Sequence
  • Betula / chemistry
  • Betula / immunology
  • Catharanthus / chemistry*
  • Catharanthus / genetics
  • Catharanthus / immunology
  • Circular Dichroism
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Protein Folding
  • Recombinant Proteins / analysis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Restriction Mapping
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Skin Tests
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Allergens
  • Antigens, Plant
  • Plant Proteins
  • Recombinant Proteins