title: Molecular cloning and characterization of hazel pollen protein (70 kD) as a luminal binding protein (BiP): a novel cross-reactive plant allergen.
abstract: BACKGROUND
  Tree pollen contains many allergens showing cross-reactivity to proteins from pollen, seeds, and fruits of different plant species. Amongst Fagales, responsible for several allergenic responses, hazel provides the best material to study pollen as well as food allergens in one species. The aim of this study was to identify and characterize the physiological function of an allergen from hazel pollen and to determine possible cross-reactivity to proteins from hazelnut.
  
  
  METHODS
  Monoclonal antibodies (mAbs) against hazel pollen crude extract were produced. On the basis of IgE binding, demonstrated by sera from patients allergic to hazel pollen, one mAb indicating the best correlation has been selected, and the putative allergen was purified by preparative gel electrophoresis. Isoforms were investigated by two-dimensional PAGE, and for molecular identification a hazel pollen cDNA library was constructed. In situ localization of the allergen during pollen development was performed by immunofluorescence labelling.
  
  
  RESULTS
  Immunological staining of crude hazel pollen extract with specific IgE and mAb revealed a 70-kD protein. Immunoblot studies with mAb showed cross-reactive proteins of 70-72 kD in different plant tissues and species. After protein purification, the IgE-binding reactivity of the allergen has been reconfirmed, and two isoforms were detected. Molecular cloning identified the allergen as a luminal binding protein (BiP) of the Hsp70 family with 88-92% sequence identity in various plants. Further immunocytological studies indicated involvement of BiP during pollen development.
  
  
  CONCLUSIONS
  Chaperons like BiP play an important role in protein synthesis and in the protection of cellular structures during stress-related processes. Because of their highly conserved protein sequences, we propose that such allergens could be responsible for at least a part of the allergenic cross-reactivity between proteins from different pollens and plant foods.
authors.affiliation: Department of Plant Cell Biology, Botany Institute, University of Bonn, Germany. gruehn@bgfa.ruhr-uni-bochum.de
authors.forename: Sabine
authors.initials: S
authors.lastname: Gruehn
authors.forename: Cenk
authors.initials: C
authors.lastname: Suphioglu
authors.forename: Robyn E
authors.initials: RE
authors.lastname: O'Hehir
authors.forename: Dieter
authors.initials: D
authors.lastname: Volkmann
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular / Q000379:methods; D031211:Corylus / Q000235:genetics / Q000276:immunology*; D003429:Cross Reactions / Q000276:immunology; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology*; D011058:Pollen / Q000235:genetics* / Q000276:immunology*; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D000485:Allergens; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D018840:HSP70 Heat-Shock Proteins; D010940:Plant Proteins; C077221:BIP protein, Arabidopsis; D007073:Immunoglobulin E