Involvement of Tyrosines at Fucose-binding Sites of Aleuria aurantia Lectin : Non-equal Response to Site-directed Mutagenesis among Five Sites

この論文にアクセスする

この論文をさがす

著者

抄録

Since the involvement of Tyr residues in the fucose-binding of <I>Aleuria aurantia</I> lectin (AAL) was proved by chemical modification using the Tyr-specific reagent tetranitromethane, site-directed mutagenesis was attempted. Since the tertiary structure of AAL was determined recently to be a six-bladed β-propeller fold, and five fucose-binding sites per subunit were found, based on positions of Tyr residues in the tertiary structure, three classes of mutants were constructed: 1) Tyr on the 2nd β-strand of each blade (β-2 mutants), 2) Tyr or Trp on the 3rd β-strand (β-3 mutants), and 3) Tyr outside of binding sites (other-Y mutants). The mutagenized cDNA was expressed in <I>Escherichia coli</I> as His-tag-AAL, and the hemagglutinating activity was assayed. Among 14 mutants, three β-2 mutants (Y26A, Y79A, and Y181A), and three β-3 mutants (Y92A, W149A, and Y241A) showed decreased activity. These mutated residues resided at Sites 1, 2, and 4, at the same locations relatively in the binding sites. Mutagenesis of Tyr or Trp at the corresponding locations in Sites 3 and 5 did not lead to a reduction in activity. Results indicate that the properties of Sites 1, 2, and 4 are different from those of Sites 3 and 5, and that the contribution of these two sites to the hemagglutination reaction was minor.

収録刊行物

  • Bioscience, biotechnology, and biochemistry

    Bioscience, biotechnology, and biochemistry 68(4), 841-847, 2004-04-23

    Japan Society for Bioscience, Biotechnology, and Agrochemistry

参考文献:  16件中 1-16件 を表示

被引用文献:  1件中 1-1件 を表示

各種コード

  • NII論文ID(NAID)
    10013143765
  • NII書誌ID(NCID)
    AA10824164
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    09168451
  • NDL 記事登録ID
    6930291
  • NDL 雑誌分類
    ZR7(科学技術--農林水産--農産) // ZR2(科学技術--生物学--生化学) // ZP1(科学技術--化学・化学工業)
  • NDL 請求記号
    Z53-G223
  • データ提供元
    CJP書誌  CJP引用  NDL  J-STAGE 
ページトップへ