Vacuolar serine protease is a major allergen of Cladosporium cladosporioides

Int Arch Allergy Immunol. 2008;146(4):277-86. doi: 10.1159/000121462. Epub 2008 Mar 21.

Abstract

Background: Cladosporium is an important allergenic fungus worldwide. We report here a major allergen of C. cladosporioides.

Methods: Major C. cladosporioides allergens were characterized by immunoblotting, N-terminal amino acid sequencing, protein purification and cDNA cloning.

Results: Seventy-four sera (38%) from 197 bronchial asthmatic patients demonstrated IgE binding against C. cladosporioides extracts. Among these 74 sera, 41 (55%) and 38 (51%) showed IgE binding against a 36- and a 20-kDa protein of C. cladosporioides, respectively. Both IgE-reacting components reacted with FUM20, a monoclonal antibody against fungal serine proteases. N-terminal amino acid sequencing results suggest that they are vacuolar serine proteases, and the 20-kDa component is possibly a degraded product of the 36-kDa allergen. A corresponding 5'-truncated 1,425-bp cDNA fragment was isolated. The mature protein after N-terminal processing starts with an N-terminal serine that is the ninth residue encoded by the 5'-truncated cDNA. The protein sequence deduced shares 69-72% sequence identity with Penicillium vacuolar serine proteases and was designated as Cla c 9. The purified 36-kDa Cla c 9 allergen showed proteolytic activity with peptide Z-Ala-Ala-Leu-pNA as substrate. IgE cross-reactivity was detected between the purified Cla c 9 and serine protease allergens from Aspergillusfumigatus and Penicillium chrysogenum.

Conclusion: We identified a vacuolar serine protease as a major allergen of C. cladosporioides (Cla c 9) and a major pan-allergen of prevalent airborne fungi. IgE cross-reactivity among these highly conserved serine protease pan-fungal allergens was also detectable.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Allergens / chemistry
  • Allergens / genetics
  • Allergens / immunology*
  • Allergens / isolation & purification
  • Amino Acid Sequence
  • Asthma / blood
  • Asthma / immunology*
  • Asthma / microbiology
  • Base Sequence
  • Cladosporium / enzymology*
  • Cladosporium / genetics
  • Cladosporium / immunology*
  • Cloning, Molecular
  • Cross Reactions
  • DNA, Complementary / genetics
  • Fungal Proteins / genetics
  • Fungal Proteins / immunology
  • Humans
  • Immunoblotting
  • Immunoglobulin E / immunology
  • Molecular Sequence Data
  • RNA, Fungal / chemistry
  • RNA, Fungal / genetics
  • Recombinant Proteins / immunology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Analysis, Protein
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / immunology*
  • Serine Endopeptidases / isolation & purification

Substances

  • Allergens
  • DNA, Complementary
  • Fungal Proteins
  • RNA, Fungal
  • Recombinant Proteins
  • Immunoglobulin E
  • Serine Endopeptidases

Associated data

  • GENBANK/EF407520