title: High-level expression in Escherichia coli and purification of recombinant plant profilins: comparison of IgE-binding capacity and allergenic activity.
abstract: Because of their structural similarity and ubiquitous distribution as actin binding proteins, plant profilins represent important cross-reactive allergens for almost 20% of patients suffering from Type I allergy to pollen and other plant products. The cDNAs coding for three birch profilin variants (Tyr44, Glu47, and Asn47), timothy grass profilin, and three tobacco profilin isoforms (ntprof1-3) were expressed at high levels in Escherichia coli as non-fusion proteins. The recombinant plant profilins were purified to homogeneity by poly (L-proline) affinity chromatography and showed comparable capacity to bind IgE-antibodies from profilin allergic patients. All recombinant plant profilins elicited dose-dependent histamine release from basophils of a profilin allergic patient and induced immediate type skin reactions. It is concluded that profilins from different plant species share IgE-epitopes and allergenic properties. Plant profilins therefore constitute a family of functional pan-allergens which may substitute each other for diagnosis and specific immunotherapy.
authors.affiliation: Institute of General and Experimental Pathology, University of Vienna, Austria.
authors.forename: S
authors.initials: S
authors.lastname: Vrtala
authors.forename: P
authors.initials: P
authors.lastname: Wiedemann
authors.forename: I
authors.initials: I
authors.lastname: Mittermann
authors.forename: H G
authors.initials: HG
authors.lastname: Eichler
authors.forename: W R
authors.initials: WR
authors.lastname: Sperr
authors.forename: P
authors.initials: P
authors.lastname: Valent
authors.forename: D
authors.initials: D
authors.lastname: Kraft
authors.forename: R
authors.initials: R
authors.lastname: Valenta
mesh_terms: D000485:Allergens / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D001666:Binding Sites, Antibody; D003001:Cloning, Molecular; D003285:Contractile Proteins; D003429:Cross Reactions; D018076:DNA, Complementary; D004926:Escherichia coli / Q000235:genetics; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D051304:Profilins; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012882:Skin Tests; D013045:Species Specificity
chemical_list: D000485:Allergens; D003285:Contractile Proteins; D018076:DNA, Complementary; D008840:Microfilament Proteins; D010940:Plant Proteins; D051304:Profilins; D011994:Recombinant Proteins; D007073:Immunoglobulin E