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- Title
Molecular cloning and characterization of a new Japanese cedar pollen allergen homologous to plant isoflavone reductase family.
- Authors
Kawamoto, S.; Fujimura, T.; Nishida, M.; Tanaka, T.; Aki, T.; Masubuchi, M.; Hayashi, T.; Suzuki, O.; Shigeta, S.; Ono, K.
- Abstract
Summary Background Japanese cedar (Cryptomeria japonica ) pollen is a major cause of seasonal pollinosis, and more than 10% of Japanese people suffer from this allergic disorder. However, only two major pollen allergens, Cry j 1 and Cry j 2, have been identified and exclusively characterized. Objective The aim of this study was to explore and identify important Japanese cedar pollen allergens other than Cry j 1 or Cry j 2. Methods C. japonica cDNA library was immunoscreened by rabbit antiserum raised against a partially purified cedar pollen allergen fraction. An isolated cDNA clone was inserted into a glutathione S-transferase (GST)-tagged Escherichia coli expression vector to obtain recombinant GST fusion protein. Non-fusion recombinant protein was purified by glutathione Sepharose affinity chromatography in conjunction with factor Xa cleavage of the GST moiety. IgE-binding ability of the recombinant protein was then evaluated by western blot analysis and enzyme-linked immunosorbent assay (ELISA). Results The cDNA encodes 306 amino acids with significant sequence similarity to those of plant isoflavone reductase-like proteins, which include a recently identified birch pollen allergen Bet v 5. Western blot analysis demonstrated that recombinant protein was recognized by cedar pollinosis patient IgE. In contrast to Bet v 5 being reported as a minor allergen, the recombinant protein exhibited 76% IgE binding frequency (19/25) against pollinosis patients. Conclusion Here we identified the third member of Japanese cedar pollen allergen homologous to isoflavone reductase. Its high IgE-binding frequency implicates that the isoflavone reductase homologue might be an additional major pollen allergen in C. japonica .
- Subjects
JAPAN; POLLEN; CRYPTOMERIA japonica; ALLERGIES
- Publication
Clinical & Experimental Allergy, 2002, Vol 32, Issue 7, p1064
- ISSN
0954-7894
- Publication type
Article
- DOI
10.1046/j.1365-2222.2002.01405.x