title: The vacuolar serine protease, a cross-reactive allergen from Cladosporium herbarum.
abstract: Subtilisin-like serine proteases make up one of the most important allergen-families regarding the number of individual allergens. Previously, fungal subtilisin-like serine proteases have been identified from Aspergillus-, Penicillium-, and Trichophyton-species having a prevalence of IgE-reactivity between 33% and 80%. Since IgE-cross-reactivity is a common phenomenon within fungal species we wanted to know whether this protein also represents an allergen in Cladosporium herbarum. Hence, a screening of a C. herbarum cDNA library was performed using the coding sequence of the Penicillium oxalicum vacuolar serine protease (Pen o 18) as hybridization probe, ending up with a full-length clone. Biochemical and immunological characterization of this clone revealed that C. herbarum vacuolar serine protease most likely is synthesized as a precursor with an N-terminal pro-enzyme sequence and represents a minor allergen (Cla h 9) with a prevalence of IgE-reactivity of 15.5%. Furthermore Cla h 9 specifically reacted with the two monoclonal antibodies FUM20 and PCM39, as do the vacuolar serine proteases from Aspergillus fumigatus and Penicillium species. Investigation of IgE-cross-reactivity between Cla h 9 and other fungal serine proteases revealed that cross-reactivity is higher between vacuolar than alkaline serine proteases. IgE-epitope mapping of Cla h 9 was done in order to test whether four Cla h 9-peptides having a high sequence homology to previously determined Pen ch 18-IgE-epitopes also harbour IgE-epitopes. Three-dimensional models of the vacuolar serine proteases from C. herbarum and Penicillium chrysogenum were generated for the three-dimensional localization of the Cla h 9- and Pen ch 18- IgE-reactive and -non-reactive peptides. Taken together a new C. herbarum allergen has been identified, which may be useful in a molecule-based approach of C. herbarum allergy-diagnosis and -therapy. Moreover, Cla h 9 represents a further member of the subtilisin-like serine protease allergen-family, which stresses the importance of these proteins with respect to fungal IgE-cross-reactivity.
authors.affiliation: Dept. of Cell-Biology, University of Salzburg, Salzburg, Austria.
authors.forename: Verena
authors.initials: V
authors.lastname: Pöll
authors.forename: Ursula
authors.initials: U
authors.lastname: Denk
authors.forename: Horng-Der
authors.initials: HD
authors.lastname: Shen
authors.forename: Raphael C
authors.initials: RC
authors.lastname: Panzani
authors.forename: Oliver
authors.initials: O
authors.lastname: Dissertori
authors.forename: Peter
authors.initials: P
authors.lastname: Lackner
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hemmer
authors.forename: Adriano
authors.initials: A
authors.lastname: Mari
authors.forename: Reto
authors.initials: R
authors.lastname: Crameri
authors.forename: Friedrich
authors.initials: F
authors.lastname: Lottspeich
authors.forename: Raphaela
authors.initials: R
authors.lastname: Rid
authors.forename: Klaus
authors.initials: K
authors.lastname: Richter
authors.forename: Michael
authors.initials: M
authors.lastname: Breitenbach
authors.forename: Birgit
authors.initials: B
authors.lastname: Simon-Nobbe
mesh_terms: D000485:Allergens / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001483:Base Sequence; D002963:Cladosporium / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D003429:Cross Reactions; D018604:Epitope Mapping; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D014617:Vacuoles / Q000201:enzymology* / Q000378:metabolism
chemical_list: D000485:Allergens; D005656:Fungal Proteins; D012697:Serine Endopeptidases