TY - JOUR
T1 - GM3, GM2 and GM1 mimics designed for biosensing: chemoenzymatic synthesis, target affinities and 900 MHz NMR analysis
AU - Pukin, A.V.
AU - Weijers, C.A.G.M.
AU - van Lagen, B.
AU - Wechselberger, R.
AU - Sun, B.
AU - Gilbert, M.
AU - Karwaski, M.F.
AU - Florack, D.E.A.
AU - Jacobs, B.C.
AU - Tio-Gillen, A.P.
AU - van Belkum, A.
AU - Endtz, H.P.
AU - Visser, G.M.
AU - Zuilhof, H.
PY - 2008
Y1 - 2008
N2 - Undec-10-enyl, undec-10-ynyl and 11-azidoundecyl glycoside analogues corresponding to the oligosaccharides of human gangliosides GM3, GM2 and GM1 were synthesized in high yields using glycosyltransferases from Campylobacter jejuni. Due to poor water solubility of the substrates, the reactions were carried out in methanol¿water media, which for the first time were shown to be compatible with the C. jejuni ¿-(2¿3)-sialyltransferase (CST-06) and ß-(1¿4)-N-acetylgalactosaminyltransferase (CJL-30). Bioequivalence of our synthetic analogues and natural gangliosides was examined by binding to Vibrio cholerae toxin and to the B subunit of Escherichia coli heat-labile enterotoxin. This bioequivalence was confirmed by binding mouse and human monoclonal antibodies to GM1 and acute phase sera containing IgM and IgG antibodies to GM1 from patients with the immune-mediated polyneuropathy Guillain¿Barré syndrome. The synthesized compounds were analyzed by 1D and 2D 900 MHz NMR spectroscopy. TOCSY and DQF-COSY experiments in combination with 13C¿1H correlation measurements (HSQC, HMBC) were carried out for primary structural characterization, and a complete assignment of all 1H and 13C chemical shifts is presented.
AB - Undec-10-enyl, undec-10-ynyl and 11-azidoundecyl glycoside analogues corresponding to the oligosaccharides of human gangliosides GM3, GM2 and GM1 were synthesized in high yields using glycosyltransferases from Campylobacter jejuni. Due to poor water solubility of the substrates, the reactions were carried out in methanol¿water media, which for the first time were shown to be compatible with the C. jejuni ¿-(2¿3)-sialyltransferase (CST-06) and ß-(1¿4)-N-acetylgalactosaminyltransferase (CJL-30). Bioequivalence of our synthetic analogues and natural gangliosides was examined by binding to Vibrio cholerae toxin and to the B subunit of Escherichia coli heat-labile enterotoxin. This bioequivalence was confirmed by binding mouse and human monoclonal antibodies to GM1 and acute phase sera containing IgM and IgG antibodies to GM1 from patients with the immune-mediated polyneuropathy Guillain¿Barré syndrome. The synthesized compounds were analyzed by 1D and 2D 900 MHz NMR spectroscopy. TOCSY and DQF-COSY experiments in combination with 13C¿1H correlation measurements (HSQC, HMBC) were carried out for primary structural characterization, and a complete assignment of all 1H and 13C chemical shifts is presented.
KW - covalently attached monolayers
KW - crystalline silicon surfaces
KW - nuclear-magnetic-resonance
KW - extremely mild attachment
KW - heat-labile enterotoxin
KW - one-pot synthesis
KW - campylobacter-jejuni
KW - cholera-toxin
KW - solid-phase
KW - guillain-barre
U2 - 10.1016/j.carres.2008.01.007
DO - 10.1016/j.carres.2008.01.007
M3 - Article
SN - 0008-6215
VL - 343
SP - 636
EP - 650
JO - Carbohydrate Research : an international journal
JF - Carbohydrate Research : an international journal
IS - 4
ER -