title: Calcium-dependent immunoglobulin E recognition of the apo- and calcium-bound form of a cross-reactive two EF-hand timothy grass pollen allergen, Phl p 7.
abstract: Type I allergy, an immunodisorder that affects almost 20% of the population worldwide, is based on the immunoglobulin E (IgE) recognition of per se innocuous antigens (allergens). Pollen from wind-pollinated plants belong to the most potent allergen sources. We report the isolation of a cDNA coding for a 8.6 kDa two EF-hand calcium binding allergen, Phl p 7, from a timothy grass (Phleum pratense) pollen expression cDNA library, using serum IgE from a grass pollen allergic patient. Sequence analysis identified Phl p 7 as a member of a recently discovered subfamily of pollen-specific calcium binding proteins. Recombinant Phl p 7 was expressed in Escherichia coli and purified to homogeneity as determined by mass spectroscopy. Approximately 10% of pollen allergic patients displayed IgE reactivity to rPhl p 7 and Phl p 7-homologous allergens present in pollens of monocotyledonic and dicotyledonic plants. Circular dichroism analysis of the calcium-bound and apo-rPhl p 7 indicated that differences in IgE recognition may be due to calcium-induced changes in the protein conformation. The fact that patients mount IgE antibodies against different protein conformations is interpreted as a footprint of a preferential sensitization against either form. The biological activity of rPhl p 7 was demonstrated by its ability to induce basophil histamine release and immediate type skin reactions in sensitized individuals. In conclusion, IgE binding to Phl p 7 represents an example for the conformation-dependent IgE recognition of an allergen. Recombinant Phl p 7 may be used for diagnosis and perhaps treatment of a group of patients who suffer from allergy to pollens of many unrelated plant species.
authors.affiliation: Department of Otorhinolaryngology, Institute of Medical and Chemical Laboratory Diagnostics, University of Vienna, Austria.
authors.forename: V
authors.initials: V
authors.lastname: Niederberger
authors.forename: B
authors.initials: B
authors.lastname: Hayek
authors.forename: S
authors.initials: S
authors.lastname: Vrtala
authors.forename: S
authors.initials: S
authors.lastname: Laffer
authors.forename: A
authors.initials: A
authors.lastname: Twardosz
authors.forename: L
authors.initials: L
authors.lastname: Vangelista
authors.forename: W R
authors.initials: WR
authors.lastname: Sperr
authors.forename: P
authors.initials: P
authors.lastname: Valent
authors.forename: H
authors.initials: H
authors.lastname: Rumpold
authors.forename: D
authors.initials: D
authors.lastname: Kraft
authors.forename: K
authors.initials: K
authors.lastname: Ehrenberger
authors.forename: R
authors.initials: R
authors.lastname: Valenta
authors.forename: S
authors.initials: S
authors.lastname: Spitzauer
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D003429:Cross Reactions; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D006636:Histamine Release; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000209:etiology; D007073:Immunoglobulin E / Q000378:metabolism*; D066298:In Vitro Techniques; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006109:Poaceae / Q000235:genetics / Q000276:immunology; D011058:Pollen / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D000485:Allergens; D001059:Apoproteins; D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D002118:Calcium