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 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-γ (IFN-γ) 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.

[1]  M. Kiso,et al.  Synthetic Studies on Sialoglycoconjugates 90: Total Synthesis of Sulfated Glucuronyl Paraglobosides , 1996 .

[2]  P. Crocker.  Mammalian Carbohydrate Recognition Systems , 2012, Results and Problems in Cell Differentiation.

[3]  M. Kiso,et al.  Synergistic interactions of the two classes of ligand, sialyl‐Lewisa/x fuco‐oligosaccharides and short sulpho‐motifs, with the P‐ and L‐selectins: implications for therapeutic inhibitor designs , 2002, Immunology.

[4]  R. U. Margolis,et al.  High prevalence of 2-mono- and 2,6-di-substituted manol-terminating sequences among O-glycans released from brain glycopeptides by reductive alkaline hydrolysis. , 1999, European journal of biochemistry.

[5]  M. Kiso,et al.  Stereocontrolled synthesis of sialyl Lewis X ceramide consisting of a pentasaccharide recognized by the selectin family. , 1992, Carbohydrate Research.

[6]  A. Surolia,et al.  Photoswitchable multivalent sugar ligands: synthesis, isomerization, and lectin binding studies of azobenzene-glycopyranoside derivatives. , 2002, Journal of the American Chemical Society.

[7]  Shaoyi Liu,et al.  Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells , 2002, Nature Biotechnology.

[8]  C. Augé,et al.  Chemoenzymatic synthesis of a 3IV,6III-disulfated Lewis(x) pentasaccharide, a candidate ligand for human L-selectin. , 1997, Carbohydrate research.

[9]  A. M. Lawson,et al.  Sialyl-Lewis(x) sequence 6-O-sulfated at N-acetylglucosamine rather than at galactose is the preferred ligand for L-selectin and de-N-acetylation of the sialic acid enhances the binding strength. , 1997, Biochemical and biophysical research communications.

[10]  T. Feizi,et al.  Carbohydrate ligands for the leukocyte-endothelium adhesion molecules, selectins. , 2001, Results and problems in cell differentiation.

[11]  A. Streit,et al.  The Lex Carbohydrate Sequence Is Recognized by Antibody to L5, a Functional Antigen in Early Neural Development , 1996, Journal of neurochemistry.

[12]  L. Federici,et al.  Carbohydrate Bioengineering Interdisciplinary approaches , 2002 .

[13]  R. Lemoine,et al.  Synthesis of 3E- And 6E-Monosulfated and 3E,6E-Disulfated Lewis X Pentasaccharides, Candidate Ligands for Human L-Selectin , 2000 .

[14]  A. Surolia,et al.  Interaction between lectin from Ricinus communis and liposomes containing gangliosides , 1975, Nature.

[15]  A. M. Lawson,et al.  Core-branching pattern and sequence analysis of mannitol-terminating oligosaccharides by neoglycolipid technology. , 1999, Analytical biochemistry.

[16]  K. Tashiro,et al.  Versican Interacts with Chemokines and Modulates Cellular Responses* , 2001, The Journal of Biological Chemistry.

[17]  R. W. Baldwin,et al.  A monoclonal antibody against human colonic adenoma recognizes difucosylated Type-2-blood-group chains , 1983, Bioscience reports.

[18]  Christopher W. Cairo,et al.  Hitting the sweet spot , 2002, Nature Biotechnology.

[19]  B. Geiger,et al.  Immunocytochemical localization of native chondroitin-sulfate in tissues and cultured cells using specific monoclonal antibody , 1984, Cell.

[20]  R. Carruthers,et al.  Optimal procedure for combined high‐performance thin‐layer chromatography/high‐sensitivity liquid secondary ion mass spectrometry , 1991 .

[21]  A. M. Lawson,et al.  Nonreductive release of O-linked oligosaccharides from mucin glycoproteins for structure/function assignments as neoglycolipids: application in the detection of novel ligands for E-selectin. , 1997, Glycobiology.

[22]  P. Sartipy,et al.  CD44, a Cell Surface Chondroitin Sulfate Proteoglycan, Mediates Binding of Interferon-γ and Some of Its Biological Effects on Human Vascular Smooth Muscle Cells* , 1999, The Journal of Biological Chemistry.

[23]  W. C. Still,et al.  Parallel Synthesis and Screening of a Solid Phase Carbohydrate Library , 1996, Science.

[24]  T. Mongini,et al.  Immunohistochemical localization of chondroitin sulfate in normal and pathological human muscle , 1986, Journal of the Neurological Sciences.

[25]  E. Hounsell,et al.  The monoclonal antibody anti‐SSEA‐1 discriminates between fucosylated type 1 and type 2 blood group chains , 1981, FEBS letters.

[26]  C. Rogers,et al.  Expression of the (1,3)Fucosyltransferase Fuc-TVII in Lymphoid Aggregate High Endothelial Venules Correlates with Expression of L-Selectin Ligands (*) , 1996, The Journal of Biological Chemistry.

[27]  A. C. Stuart,et al.  Novel sulfated ligands for the cell adhesion molecule E-selectin revealed by the neoglycolipid technology among O-linked oligosaccharides on an ovarian cystadenoma glycoprotein. , 1992, Biochemistry.

[28]  M. Kiso,et al.  L-selectin Interactions with Novel Mono- and Multisulfated Lewisx Sequences in Comparison with the Potent Ligand 3′-Sulfated Lewisa * , 1999, The Journal of Biological Chemistry.

[29]  Y. Lee,et al.  Novel approach to the study of the antigenicities and receptor functions of carbohydrate chains of glycoproteins. , 1985, Biochemical and biophysical research communications.

[30]  M. Kiso,et al.  Synthesis and biological activities of three sulfated sialyl Le(x) ganglioside analogues for clarifying the real carbohydrate ligand structure of L-selectin. , 1996, Bioorganic & medicinal chemistry.

[31]  P. Crocker,et al.  Siglecs in the immune system , 2001, Immunology.

[32]  T. Mizuochi,et al.  Improved procedure for the construction of neoglycolipids having antigenic and lectin-binding activities, from reducing oligosaccharides. , 1988, The Biochemical journal.

[33]  A. Surolia,et al.  Synthetic glycolipids: interaction with galactose-binding lectin and hepatic cells. , 1981, Archives of biochemistry and biophysics.

[34]  A. M. Lawson,et al.  Fluorescent neoglycolipids. Improved probes for oligosaccharide ligand discovery. , 2000, European journal of biochemistry.

[35]  A Lubineau,et al.  First synthesis of the 3'-sulfated Lewis(a) pentasaccharide, the most potent human E-selectin ligand so far. , 1994, Bioorganic & medicinal chemistry.

[36]  D. Solter,et al.  Stage-specific embryonic antigen involves αl→ 3 fucosylated type 2 blood group chains , 1981, Nature.

[37]  Milan Mrksich,et al.  Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. , 2002, Chemistry & biology.

[38]  J. Raynes,et al.  Human serum amyloid P is a multispecific adhesive protein whose ligands include 6‐phosphorylated mannose and the 3‐sulphated saccharides galactose, N‐acetylgalactosamine and glucuronic acid. , 1992, The EMBO journal.

[39]  R. U. Margolis,et al.  Brain Contains HNK-1 Immunoreactive O-Glycans of the Sulfoglucuronyl Lactosamine Series that Terminate in 2-Linked or 2,6-Linked Hexose (Mannose)* , 1997, The Journal of Biological Chemistry.

[40]  A. M. Lawson,et al.  Specificity of mild periodate oxidation of oligosaccharide-alditols: relevance to the analysis of the core-branching pattern of O-linked glycoprotein oligosaccharides. , 1993, Carbohydrate research.

[41]  A. M. Lawson,et al.  Microscale sequencing of O-linked oligosaccharides using mild periodate oxidation of alditols, coupling to phospholipid and TLC-MS analysis of the resulting neoglycolipids. , 1990, European journal of biochemistry.

[42]  J. Evans,et al.  Structure of sulfated glucuronyl glycolipids in the nervous system reacting with HNK-1 antibody and some IgM paraproteins in neuropathy. , 1986, The Journal of biological chemistry.

[43]  A. Helenius,et al.  Intracellular functions of N-linked glycans. , 2001, Science.

[44]  M. Kiso,et al.  Valency dependent patterns of binding of human L-selectin toward sialyl and sulfated oligosaccharides of Le(a) and Le(x) types: relevance to anti-adhesion therapeutics. , 1997, Biochemistry.

[45]  K. Karlsson.  Meaning and therapeutic potential of microbial recognition of host glycoconjugates , 1998, Molecular microbiology.

[46]  A. M. Lawson,et al.  Neoglycolipids: probes of oligosaccharide structure, antigenicity, and function. , 1994, Methods in Enzymology.

[47]  M. Bednarski,et al.  Direct colorimetric detection of a receptor-ligand interaction by a polymerized bilayer assembly. , 1993, Science.

[48]  P. Crocker,et al.  Carbohydrate recognition systems: functional triads in cell-cell interactions. , 1996, Current opinion in structural biology.

[49]  A. M. Lawson,et al.  10E4 Antigen of Scrapie Lesions Contains an Unusual Nonsulfated Heparan Motif* , 2001, The Journal of Biological Chemistry.

[50]  T. Feizi.  Progress in deciphering the information content of the ‘glycome’ – a crescendo in the closing years of the millennium , 2000, Glycoconjugate Journal.