Analysis of the three-dimensional antigenic structure of giant ragweed allergen, Amb t 5

Thorunn Rafnar, Mary Elizabeth Brummet, Donna Bassolino-Klimas, William J. Metzler, David G. Marsh

Research output: Contribution to journalArticle

Abstract

The ragweed allergens Amb t 5 and Amb a 5 are among the smallest inhaled protein allergens known, containing a single, immunodominant T-cell epitope. In this study we analyzed the B-cell epitope structure of Amb t 5. The three- dimensional structures of Amb t 5 and Amb a 5 have been determined by NMR spectroscopy, providing a rare opportunity to analyze three-dimensional antigenic sites. Amb t 5 residues likely to be important for antigenicity were identified by examining the surface area of Amb t 5 accessible to a probe of the size of an antibody molecule. After changing these residues to the corresponding Amb a 5 residues, recombinant proteins were purified and tested for loss of antigenic activity. Inhibition radio-immunoassays, using sera from 8 individuals who had received immunotherapy with giant ragweed extract, allowed the mutations to be divided into three groups: (1). mutations that had little or no effect on antibody binding, (2) mutations that caused a loss of antigenic activity to a different degree in different sera and (3) mutations that drastically reduced antigenic activity in all sera tested. This last set of mutations clustered in the third loop of Amb t 5, suggesting that antibody recognition of Amb t 5, like T-cell recognition, is primarily directed towards a single, immunodominant site.

Original languageEnglish (US)
Pages (from-to)459-467
Number of pages9
JournalMolecular Immunology
Volume35
Issue number8
DOIs
StatePublished - May 1 1998
Externally publishedYes

Fingerprint

Ambrosia
Allergens
Mutation
Immunodominant Epitopes
Antibodies
Serum
B-Lymphocyte Epitopes
T-Lymphocyte Epitopes
Radio
Immunoassay
Recombinant Proteins
Immunotherapy
Magnetic Resonance Spectroscopy
T-Lymphocytes
Proteins

Keywords

  • Allergens
  • Epitope structure
  • Ragweed

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

Analysis of the three-dimensional antigenic structure of giant ragweed allergen, Amb t 5. / Rafnar, Thorunn; Brummet, Mary Elizabeth; Bassolino-Klimas, Donna; Metzler, William J.; Marsh, David G.

In: Molecular Immunology, Vol. 35, No. 8, 01.05.1998, p. 459-467.

Research output: Contribution to journalArticle

Rafnar, Thorunn ; Brummet, Mary Elizabeth ; Bassolino-Klimas, Donna ; Metzler, William J. ; Marsh, David G. / Analysis of the three-dimensional antigenic structure of giant ragweed allergen, Amb t 5. In: Molecular Immunology. 1998 ; Vol. 35, No. 8. pp. 459-467.
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