Enzymatic synthesis of spacer-linked divalent glycosides carrying N-acetylglucosamine and N-acetyllactosamine: analysis of cross-linking activities with WGA.
Divalent glycosides carrying N-acetyl-d-glucosamine (GlcNAc) and N-acetyllactosamine (LacNAc) were designed and prepared as glycomimetics. First, hexan-1,6-diyl bis-(2-acetamido-2-deoxy-beta-d-glucopyranoside) (GlcNAc-Hx-GlcNAc) and 3,6-dioxaoct-1,8-diyl bis-(2-acetamido-2-deoxy-beta-d-glucopyranoside) (GlcNAc-Doo-GlcNAc) were enzymatically synthesized by transglycosylation of an N,N'N'',N'''-tetraacetylchitotetraose [(GlcNAc)(4)] donor with a primary diol acceptor, utilizing a chitinolytic enzyme from Amycolatopsis orientalis. The resulting divalent glycosides were further converted to the respective hexan-1,6-diyl bis-[beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside] (LacNAc-Hx-LacNAc) and 6-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-hexyl beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside (LacNAc-Hx-GlcNAc), and respective 3,6-dioxaoct-1,8-diyl bis-[beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside] (LacNAc-Doo-LacNAc) and 8-(2-acetamido-2-deoxy-beta-d-glucopyranosyl)-3,6-dioxaoctyl beta-d-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-d-glucopyranoside (LacNAc-Doo-GlcNAc) by galactosyltransferase. The interaction of wheat germ agglutinin (WGA) with a series of divalent glycosides and related compounds were studied using a biosensor based on surface plasmon resonance (SPR) and by precipitation analysis. Our results demonstrated that divalent glycosides carrying GlcNAc on both sides and GlcNAc and LacNAc on each side are capable of precipitating WGA as divalent ligands, but that the corresponding monovalent controls and divalent glycosides carrying LacNAc on both sides are unable to precipitate the lectin and bind as univalent ligands.
Misawa
Yoshinori
Y
Science of Biological Resource, The United Graduate School of Agricultural Science, Gifu University, Yanagido, Gifu 501-1193, Japan.
Akimoto
Takashi
T
Amarume
Satoshi
S
Murata
Takeomi
T
Usui
Taichi
T
D000117:Acetylglucosamine / Q000737:chemistry*; D000192:Actinomycetales / Q000201:enzymology; D000606:Amino Sugars / Q000737:chemistry*; D011232:Chemical Precipitation; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000378:metabolism; D005700:Galactosyltransferases / Q000378:metabolism; D006027:Glycosides / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D008024:Ligands; D020349:Surface Plasmon Resonance; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
D000606:Amino Sugars; D003432:Cross-Linking Reagents; D006027:Glycosides; D008024:Ligands; D014909:Wheat Germ Agglutinins; C000458:N-acetyllactosamine; D005700:Galactosyltransferases; D000117:Acetylglucosamine