- 作者: Wen-Chia Wu Ming F. Tam Ho-Jen Peng Lai-Chin Tsai Chin-Wen Chi Zo-Nan Chang
- 作者服務機構: Institute of Biotechnology in Medicine, National Yang-Ming University, Insitute of Molecular Biology, Academia Sinica, and Department of Medical research and Education, Veterans General Hospital-Taipei, Taipei, Taiwan, ROC
- 中文摘要: --
- 英文摘要: Cyn d Bd46K, a 46-kD component of Bermuda grass (Cy-nodon dactylon) pollen, had been identified as an aller-genic constituent. In the present study two-dimensional(2D) gel electrophoresis illustrated the presence of fiveacidic isoforms in Cyn d Bd46K, and this molecule waspurified by monoclonal antibody (MAb) affinity chroma-tography for further characterization. Using a digoxige-nin-labeled lectin-binding assay, the elucidating proteinwas disclosed to be a glycoprotein with terminal man-nose. The involvement of a carbohydrate moiety in theallergenicity and antigenicity of the elucidated moleculewas demonstrated with sodium-periodate-treated Cyn dBd46K, which reduced binding to its specific MAb andhuman IgE. We were unable to identify the N-terminalamino acid sequences of Cyn d Bd46K, but some internalamino acid sequences were disclosed by microsequenc-ing some fragments cleaved by Achromobacter prote-ase I and fractionated by reversed-phase column chro-matography. The amino acid sequences of 4 identifiedCyn d Bd46K internal peptide fragments were found tobe 25-71% identical with that of cytochrome c oxidase IIIfrom corn grass pollen.The present study providedimportant information for future experiments on the mo-lecular cloning of the elucidated allergen.
- 中文關鍵字: --
- 英文關鍵字: Bermuda grass pollen. Cyn d Bd46K. Monoclonal antibody. Affinity chromatography. Internal sequencing