Glycosyltransferase Microarray Displayed on the Glycolipid LB Membrane

Authors

  • Noriko Nagahori,

    1. Laboratory for Bio-Macromolecular Chemistry Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10 W8, Kita-ku, Sapporo 060–0810, Japan Fax: (+81)-11-706-3435
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  • Kenichi Niikura,

    1. Sapporo Laboratory for Glycocluster Project, Japan Bioindustry Association, N10 W8, Kita-ku, Sapporo 060–0810, Japan
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  • Reiko Sadamoto,

    1. Sapporo Laboratory for Glycocluster Project, Japan Bioindustry Association, N10 W8, Kita-ku, Sapporo 060–0810, Japan
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  • Masahiro Taniguchi,

    1. Laboratory for Bio-Macromolecular Chemistry Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10 W8, Kita-ku, Sapporo 060–0810, Japan Fax: (+81)-11-706-3435
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  • Akihiko Yamagishi,

    1. Laboratory for Bio-Macromolecular Chemistry Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10 W8, Kita-ku, Sapporo 060–0810, Japan Fax: (+81)-11-706-3435
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  • Kenji Monde,

    1. Laboratory for Bio-Macromolecular Chemistry Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10 W8, Kita-ku, Sapporo 060–0810, Japan Fax: (+81)-11-706-3435
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  • Shin-Ichiro Nishimura

    1. Laboratory for Bio-Macromolecular Chemistry Division of Biological Sciences, Graduate School of Science, Hokkaido University, N10 W8, Kita-ku, Sapporo 060–0810, Japan Fax: (+81)-11-706-3435
    2. Sapporo Laboratory for Glycocluster Project, Japan Bioindustry Association, N10 W8, Kita-ku, Sapporo 060–0810, Japan
    3. Glycochemosynthesis Team, Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo 062–8517, Japan
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Abstract

β(1→4) Galactosyltransferase expressed as a fusion protein with maltose binding protein (MBP-GalT) was displayed specifically on a Langmuir–Blodgett (LB) membrane prepared by photopolymerization of maltotriose-carrying glycolipid (1) with 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3-phosphocholine (2). The catalytic activity of MBP-GalT on the LB film was directly monitored by the surface plasmon resonance (SPR) method using a GlcNAc-carrying water-soluble polymer (3) as an acceptor substrate. Highly sensitive sigmoidal-type signals were obtained upon the addition of the acceptor substrate in the presence of the donor substrate, UDP-galactose (UDP-Gal), while the binding of 3 was not detected in the absence of UDP-Gal. The intensities of the signals were dependent on the amount of immobilized MBP-GalT on the LB film, which was estimated from the images obtained by atomic force microscope (AFM).

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