Molecular Design of Fluorescent Labeled Glycosides as Acceptor Substrates for Sialyltransferases

• OGATA Makoto

  Department of Bioscience, Graduate School of Science and Technology, Shizuoka University

• OBARA Takakiyo

  Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University

• CHUMA Yasushi

  Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University

• MURATA Takeomi

  Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University

• PARK Enoch Y.

  Department of Bioscience, Graduate School of Science and Technology, Shizuoka University

• USUI Taichi

  Department of Bioscience, Graduate School of Science and Technology, Shizuoka University Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University

Abstract

A series of dansyl-labeled glycosides with di-, tetra-, and hexasaccharides carrying the terminal N-acetyllactosamine (LacNAc) sequence were synthesized as acceptor substrates for α2,6- and α2,3-sialyltransferases. As an alternative design, dansyl-labeled LacNAc glycoside carrying a long-spacer linked glycan was engineered by replacement of the LacNAc or lactose units with an alkyl chain. In addition, we designed a dansyl-labeled bi-antennary LacNAc glycoside as an N-linked oligosaccharide mimetic, such as asialo-α₁-acid glycoprotein. The kinetic parameters for the transfer reaction of synthesized dansyl-labeled glycosides by sialyltransferases were determined by the fluorescent HPLC method. The catalytic efficiencies (V_max/K_m) of acceptor substrates carrying the terminal LacNAc sequence with various length glycans in the array for α2,6- and α2,3-sialyltransferases decreased in a glycan length-dependent manner. Furthermore, of the acceptor substrates tested, dansyl-labeled bi-antennary LacNAc glycoside displayed the most favorable K_m value for α2,6- and α2,3-sialyltransferases.

Journal

• Bioscience, Biotechnology, and Biochemistry

  Bioscience, Biotechnology, and Biochemistry 74 (11), 2287-2292, 2010

  Japan Society for Bioscience, Biotechnology, and Agrochemistry

Keywords

• sialyltransferase
• enzyme assay
• chemoenzymatic synthesis
• substrate specificity
• bi-antennary oligosaccharide

Details

• CRID: 1390001206479384832
• NII Article ID: 10027561679
• NII Book ID: AA10824164
• DOI: 10.1271/bbb.100505
• ISSN: 13476947
• ISSN: 09168451
• NDL BIB ID: 10899892
• Web Site: http://id.ndl.go.jp/bib/10899892
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I10899892
• Web Site: http://www.tandfonline.com/doi/pdf/10.1271/bbb.100505
• Text Lang: en
• Data Source:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• Abstract License Flag: Disallowed