Molecular characterization, expression in Escherichia coli, and epitope analysis of a two EF-hand calcium-binding birch pollen allergen, Bet v 4

Anna Twardosz, Brigitte Hayek, Susanne Seiberler, Luca Vangelista, Lena Elfman, Hans Grönlund, Dietrich Kraft, Rudolf Valenta

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Birch pollen belongs to the most potent elicitors of Type I allergic reactions in early spring. Using serum IgE from a birch pollen allergic patient, two cDNA clones (clone 6 and clone 13) were isolated from a birch pollen expression cDNA library constructed in phage λgt11. Clone 6 encoded a 9.3 kD two EF-hand calcium-binding protein, designated Bet v 4, with significant end to end sequence homology to EF-hand calcium-binding allergens from weed and grass pollen. Recombinant Bet v 4, expressed as β-galactosidase fusion protein, reacted with serum IgE from approximately 20% of pollen allergic individuals. Depletion of allergenbound calcium by EGTA treatment lead to a substantial reduction of IgE-binding to Bet v 4, indicating that protein-bound calcium is necessary for the maintenance of IgE-epitopes. The greatly reduced IgE-binding capacity of clone 13, a Bet v 4 fragment that lacked the 16 N-terminal amino acids, indicated that the N-terminus contributes significantly to the proteins IgE-binding capacity. By IgE-inhibition experiments it was demonstrated that recombinant Bet v 4 shared IgE-epitopes with natural Bet v 4 and a homologous timothy grass pollen allergen. Recombinant Bet v 4 may therefore be considered as a relevant crossreactive plant allergen, which may be used for diagnosis and treatment of patients suffering from multivalent plant allergies.

Original languageEnglish
Pages (from-to)197-204
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume239
Issue number1
DOIs
Publication statusPublished - Oct 9 1997
Externally publishedYes

Fingerprint

EF Hand Motifs
Betula
Pollen
Allergens
Escherichia coli
Immunoglobulin E
Epitopes
Calcium
Clone Cells
Hypersensitivity
Phleum
Galactosidases
Allergies
Galectin 3
Bacteriophages
Calcium-Binding Proteins
Egtazic Acid
Sequence Homology
Poaceae
Serum

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Molecular characterization, expression in Escherichia coli, and epitope analysis of a two EF-hand calcium-binding birch pollen allergen, Bet v 4. / Twardosz, Anna; Hayek, Brigitte; Seiberler, Susanne; Vangelista, Luca; Elfman, Lena; Grönlund, Hans; Kraft, Dietrich; Valenta, Rudolf.

In: Biochemical and Biophysical Research Communications, Vol. 239, No. 1, 09.10.1997, p. 197-204.

Research output: Contribution to journalArticle

Twardosz, Anna ; Hayek, Brigitte ; Seiberler, Susanne ; Vangelista, Luca ; Elfman, Lena ; Grönlund, Hans ; Kraft, Dietrich ; Valenta, Rudolf. / Molecular characterization, expression in Escherichia coli, and epitope analysis of a two EF-hand calcium-binding birch pollen allergen, Bet v 4. In: Biochemical and Biophysical Research Communications. 1997 ; Vol. 239, No. 1. pp. 197-204.
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