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Isolation and partial characterization of a 46-kD allergen of Bermuda grass pollen

  • Original Paper
  • Published:
Journal of Biomedical Science

Abstract

Cyn d Bd46K, a 46-kD component of Bermuda grass(Cynodon dactylon) pollen, had been identified as an allergenic constituent. In the present study two-dimensional (2D) gel electrophoresis illustrated the presence of five acidic isoforms in Cyn d Bd46K, and this molecule was purified by monoclonal antibody (MAb) affinity chromatography for further characterization. Using a digoxigenin-labeled lectin-binding assay, the elucidating protein was disclosed to be a glycoprotein with terminal mannose. The involvement of a carbohydrate moiety in the allergenicity and antigenicity of the elucidated molecule was demonstrated with sodium-periodate-treated Cyn d Bd46K, which reduced binding to its specific MAb and human IgE. We were unable to identify the N-terminal amino acid sequences of Cyn d Bd46K, but some internal amino acid sequences were disclosed by microsequencing some fragments cleaved byAchromobacter protease I and fractionated by reversed-phase column chromatography. The amino acid sequences of 4 identified Cyn d Bd46K internal peptide fragments were found to be 25–71% identical with that of cytochrome c oxidase III from corn grass pollen. The present study provided important information for future experiments on the molecular cloning of the elucidated allergen.

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Authors and Affiliations

  1. Institute of Biotechnology in Medicine, National Yang-Ming University, Taiwan, ROC

    Wen-Chia Wu & Zo-Nan Chang

  2. Institute of Molecular Biology, Academic Sinica, Taiwan, ROC

    Ming F. Tam

  3. Department of Medical Research and Education, Veterans General Hospital-Taipei, Taipei, Taiwan, ROC

    Ho-Jen Peng, Lai-Chin Tsai & Chin-Wen Chi

  4. Faculty of Medical Technology, National Yang-Ming University, Taipei, Taiwan 112 (ROC)

    Zo-Nan Chang

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  1. Wen-Chia Wu
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  2. Ming F. Tam
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  4. Lai-Chin Tsai
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  5. Chin-Wen Chi
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Wu, WC., Tam, M.F., Peng, HJ. et al. Isolation and partial characterization of a 46-kD allergen of Bermuda grass pollen. J Biomed Sci 8, 342–348 (2001). https://doi.org/10.1007/BF02258376

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  • DOI: https://doi.org/10.1007/BF02258376

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