Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions.
We describe microarrays of oligosaccharides as neoglycolipids and their robust display on nitrocellulose. The arrays are obtained 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 Lewis(x) sequence based on N-acetyllactosamine recognized by anti-L5, and a paucity of the Lewis(x) 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-gamma (IFN-gamma) and the chemokine RANTES to sulfated sequences such as HNK-1, sulfo-Lewis(x), and sulfo-Lewis(a), 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.
Fukui
Shigeyuki
S
Glycosciences Laboratory, Imperial College Faculty of Medicine, Northwick Park Hospital, Watford Road, Harrow, Middlesex HA1 3UJ, UK.
Feizi
Ten
T
Galustian
Christine
C
Lawson
Alexander M
AM
Chai
Wengang
W
D000818:Animals; D001921:Brain / Q000378:metabolism; D002241:Carbohydrates / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D003101:Collodion; D006017:Glycolipids / Q000737:chemistry*; D015335:Molecular Probes / Q000138:chemical synthesis / Q000378:metabolism; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000145:classification; D011134:Polysaccharides / Q000032:analysis / Q000378:metabolism; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D011817:Rabbits; D011956:Receptors, Cell Surface; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002241:Carbohydrates; D002352:Carrier Proteins; D006017:Glycolipids; D015335:Molecular Probes; D009844:Oligosaccharides; D011134:Polysaccharides; D011506:Proteins; D011956:Receptors, Cell Surface; C055213:saccharide-binding proteins; D003101:Collodion