title: Allergen motifs and the prediction of allergenicity.
abstract: We have recently shown that the majority of allergens can be represented by allergen motifs. This observation prompted us to experimentally investigate the synthesized peptides corresponding to the in silico motifs with regard to potential IgE binding and cross-reactions with allergens. Two motifs were selected as examples to conduct in vitro studies. From the first motif, derived from allergenic MnSOD sequences, the motif stretch of the allergen Asp f 6 was selected and synthesized as a peptide (MnSOD Mot). The corresponding full-length MnSOD was also expressed in Escherichia coli and both were compared for IgE reactivity with sera of patients reacting to the MnSOD of Aspergillus fumigatus or Malassezia sympodialis. For the second motif, the invertebrate tropomyosin sequences were aligned and a motif consensus sequence was expressed as a recombinant protein (Trop Mot). The IgE reactivity of Trop Mot was analyzed in ELISA and compared to that of recombinant tropomyosin from the shrimp Penaeus aztecus (rPen a 1) in ImmunoCAP. MnSOD Mot was weakly recognized by some of the tested sera, suggesting that the IgE binding epitopes of a multimeric globular protein such as MnSOD cannot be fully represented by a motif peptide. In contrast, the motif Trop Mot showed the same IgE reactivity as shrimp full-length tropomyosin, indicating that the major allergenic reactivity of a repetitive structure such as tropomyosin can be covered by a motif peptide. Our results suggest that the motif-generating algorithm may be used for identifying major IgE binding structures of coiled-coil proteins.
authors.affiliation: Institute of Immunology, Inselspital, 3010 Bern, Switzerland.
authors.forename: Pamela
authors.initials: P
authors.lastname: Marti
authors.forename: Renato
authors.initials: R
authors.lastname: Truffer
authors.forename: Michael B
authors.initials: MB
authors.lastname: Stadler
authors.forename: Elsbeth
authors.initials: E
authors.lastname: Keller-Gautschi
authors.forename: Reto
authors.initials: R
authors.lastname: Crameri
authors.forename: Adriano
authors.initials: A
authors.lastname: Mari
authors.forename: Peter
authors.initials: P
authors.lastname: Schmid-Grendelmeier
authors.forename: Sylvia M
authors.initials: SM
authors.lastname: Miescher
authors.forename: Beda M
authors.initials: BM
authors.lastname: Stadler
authors.forename: Monique
authors.initials: M
authors.lastname: Vogel
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant; D001483:Base Sequence; D003429:Cross Reactions; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000276:immunology; D016415:Sequence Alignment; D012758:Shellfish; D013329:Structure-Activity Relationship; D013482:Superoxide Dismutase / Q000235:genetics / Q000276:immunology*; D014335:Tropomyosin / Q000235:genetics / Q000276:immunology*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D014335:Tropomyosin; D007073:Immunoglobulin E; C115317:Asp f 6 allergen; D013482:Superoxide Dismutase