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Glycosyltransferase Microarray Displayed on the Glycolipid LB Membrane
Author(s) -
Nagahori Noriko,
Niikura Kenichi,
Sadamoto Reiko,
Taniguchi Masahiro,
Yamagishi Akihiko,
Monde Kenji,
Nishimura ShinIchiro
Publication year - 2003
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.200202204
Subject(s) - chemistry , maltotriose , glycolipid , substrate (aquarium) , galactosyltransferase , surface plasmon resonance , membrane , maltose , acceptor , galactose , biochemistry , enzyme , nanoparticle , nanotechnology , oceanography , materials science , physics , condensed matter physics , geology
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).