title: Vacuolar serine protease is a major allergen of Cladosporium cladosporioides.
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.
  
  
  CONCLUSIONS
  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.
authors: Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan.
authors: Hong
authors: H
authors: Chou
authors: Ming F
authors: MF
authors: Tam
authors: Lin-Hau
authors: LH
authors: Lee
authors: Chi-Huei
authors: CH
authors: Chiang
authors: Hsiao-Yun
authors: HY
authors: Tai
authors: Raphael C
authors: RC
authors: Panzani
authors: Horng-Der
authors: HD
authors: Shen
mesh_terms: D000328:Adult; D000485:Allergens / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001249:Asthma / Q000097:blood / Q000276:immunology* / Q000382:microbiology; D001483:Base Sequence; D002963:Cladosporium / Q000201:enzymology* / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D003429:Cross Reactions; D018076:DNA, Complementary / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000485:Allergens; D018076:DNA, Complementary; D005656:Fungal Proteins; D012331:RNA, Fungal; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D012697:Serine Endopeptidases