Mapping of IgE-binding epitopes on the recombinant major group I allergen of velvet grass pollen, rHol l 1

J Allergy Clin Immunol. 1997 Jun;99(6 Pt 1):781-7. doi: 10.1016/s0091-6749(97)80012-7.


Background: New and more successful approaches to diagnosis and therapy of allergic diseases require a more subtle understanding of the structure and the epitopes on the allergen molecule.

Objective: This study was done to obtain more information on the structure and the IgE-binding epitopes of a major allergen of velvet grass pollen, Hol l 1.

Methods: We cloned Hol l 1 from a complementary DNA library and performed B-cell epitope mapping with 21 recombinant fragments expressed as fusion proteins in Escherichia coli. The fragments were analyzed by Western blotting with sera from 50 different patients.

Results: The patients' sera individually recognized at least four different IgE-binding regions (amino acids 1 to 27, 61 to 76, 84 to 105, and 158 to 240). According to their binding patterns with these epitopes, they were divided into five groups. Most sera (92%) bound to the C-terminal peptide (158 to 240), which consists of more than 80 amino acids, whereas there was virtually no binding to smaller fragments covering this region. In contrast to the C-terminal peptide, the IgE-binding peptides on the N terminus and on the middle region of the molecule were of a smaller size (15 to 30 amino acids).

Conclusions: The major group I allergen of velvet grass bears at least four different IgE-binding epitopes, which were individually recognized by sera from different patients. The C terminus represents the major IgE-binding region and contains at least one discontinuous IgE-binding epitope, whereas the N terminus and middle region of Hol l 1 seem to contain continuous IgE-binding epitopes.

MeSH terms

  • Allergens / biosynthesis
  • Allergens / chemistry*
  • Allergens / genetics
  • Amino Acid Sequence
  • Antigens, Plant
  • Base Sequence
  • Binding Sites, Antibody*
  • DNA, Complementary / isolation & purification
  • Epitope Mapping*
  • Epitopes, B-Lymphocyte / chemistry
  • Glycoproteins / biosynthesis
  • Glycoproteins / genetics
  • Glycoproteins / immunology*
  • Humans
  • Immunoglobulin E / metabolism*
  • Molecular Sequence Data
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Plant Proteins / biosynthesis
  • Plant Proteins / genetics
  • Plant Proteins / immunology*
  • Poaceae / chemistry
  • Poaceae / immunology
  • Pollen / chemistry*
  • Pollen / immunology
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology*


  • Allergens
  • Antigens, Plant
  • DNA, Complementary
  • Epitopes, B-Lymphocyte
  • Glycoproteins
  • HOL L 1 protein, Holcus lanatus
  • Peptide Fragments
  • Plant Proteins
  • Recombinant Proteins
  • Immunoglobulin E

Associated data

  • GENBANK/Z27084
  • GENBANK/Z68893