The vacuolar serine protease, a cross-reactive allergen from Cladosporium herbarum

Mol Immunol. 2009 Apr;46(7):1360-73. doi: 10.1016/j.molimm.2008.11.017. Epub 2009 Jan 21.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / genetics
  • Allergens / immunology*
  • Allergens / isolation & purification
  • Amino Acid Sequence
  • Base Sequence
  • Cladosporium / enzymology
  • Cladosporium / genetics
  • Cladosporium / immunology*
  • Cloning, Molecular
  • Cross Reactions
  • Epitope Mapping
  • Fungal Proteins / genetics
  • Fungal Proteins / immunology
  • Fungal Proteins / isolation & purification
  • Models, Molecular
  • Molecular Sequence Data
  • Sequence Analysis, Protein
  • Sequence Homology, Amino Acid
  • Serine Endopeptidases / genetics*
  • Serine Endopeptidases / immunology*
  • Serine Endopeptidases / isolation & purification
  • Serine Endopeptidases / metabolism
  • Vacuoles / enzymology*
  • Vacuoles / metabolism


  • Allergens
  • Fungal Proteins
  • Serine Endopeptidases