Nature Biotechnology
20, 1011 - 1017 (2002)
Published online: 3 September 2002; | doi:10.1038/nbt735
Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactionsShigeyuki Fukui1, 2, Ten Feizi1, Christine Galustian1, Alexander M. Lawson1
& Wengang Chai11
Glycosciences Laboratory, Imperial College Faculty of Medicine, Northwick Park Hospital, Watford Road, Harrow, Middlesex HA1 3UJ, UK. 2
Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Kyoto, Japan (on leave).
Correspondence should be addressed to Ten Feizi t.feizi@ic.ac.ukWe describe microarrays of oligosaccharides as neoglycolipids and their robust display on nitrocellulose. The arrays are sourced from glycoproteins, glycolipids, proteoglycans, polysaccharides, whole organs, or from chemically synthesized oligosaccharides. We show that carbohydrate-recognizing proteins single out their ligands not only in arrays of homogeneous oligosaccharides but also in arrays of heterogeneous oligosaccharides. Initial applications have revealed new findings, including: (i) among O-glycans in brain, a relative abundance of the Lewisx sequence based on N-acetyllactosamine recognized by anti-L5, and a paucity of the Lewisx sequence based on poly-N-acetyllactosamine recognized by anti-SSEA-1; (ii) insights into chondroitin sulfate oligosaccharides recognized by an antiserum and an antibody (CS-56) to chondroitin sulfates; and (iii) binding of the cytokine interferon- (IFN-) and the chemokine RANTES to sulfated sequences such as HNK-1, sulfo-Lewisx, and sulfo-Lewisa, in addition to glycosaminoglycans. The approach opens the way for discovering new carbohydrate-recognizing proteins in the proteome and for mapping the repertoire of carbohydrate recognition structures in the glycome.
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