Neoglycolipid probes prepared via oxime ligation for microarray analysis of oligosaccharide-protein interactions.
Neoglycolipid technology is the basis of a microarray platform for assigning oligosaccharide ligands for carbohydrate-binding proteins. The strategy for generating the neoglycolipid probes by reductive amination results in ring opening of the core monosaccharides. This often limits applicability to short-chain saccharides, although the majority of recognition motifs are satisfactorily presented with neoglycolipids of longer oligosaccharides. Here, we describe neoglycolipids prepared by oxime ligation. We provide evidence from NMR studies that a significant proportion of the oxime-linked core monosaccharide is in the ring-closed form, and this form selectively interacts with a carbohydrate-binding protein. By microarray analyses we demonstrate the effective presentation with oxime-linked neoglycolipids of (1) Lewis(x) trisaccharide to antibodies to Lewis(x), (2) sialyllactose analogs to the sialic acid-binding receptors, siglecs, and (3) N-glycans to a plant lectin that requires an intact N-acetylglucosamine core.
Liu
Yan
Y
Glycosciences Laboratory, Imperial College London, Northwick Park and St. Mark's Campus, Harrow, Middlesex HA1 3UJ, United Kingdom.
Feizi
Ten
T
Campanero-Rhodes
María A
MA
Childs
Robert A
RA
Zhang
Yibing
Y
Mulloy
Barbara
B
Evans
Philip G
PG
Osborn
Helen M I
HM
Otto
Diana
D
Crocker
Paul R
PR
Chai
Wengang
W
D002240:Carbohydrate Sequence; D006017:Glycolipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D015335:Molecular Probes; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D010091:Oximes / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D006017:Glycolipids; D015335:Molecular Probes; D009844:Oligosaccharides; D010091:Oximes; D011506:Proteins