Abstract
In this chapter, we describe the key steps of the “designer” oligosaccharide microarray approach we followed to prove the carbohydrate binding activity and define the oligosaccharide ligands for Dectin-1, an atypical C-type lectin-like signaling receptor of the mammalian innate immune system with a key role in anti-fungal immunity. The term “designer” microarray, which we introduced in the course of the Dectin-1 study refers to a microarray of oligosaccharide probes generated from ligand-bearing glycoconjugates to reveal the oligosaccharide ligands they harbor, so that these can be isolated and characterized. Oligosaccharide probes were generated from two polysaccharides, one that was bound by Dectin-1 and known to be rich in β1,3-glucose sequence and another that was not bound and was rich in β1,6-glucose sequence and served as a negative control. The approach involved: classic ELISA-type binding assays to select the polysaccharides; partial depolymerization of the polysaccharides by chemical hydrolysis; fractionation by size of the glucan oligosaccharides obtained and determination of their chain lengths by mass spectrometry; detection of Dectin-1 ligand-positive and ligand-negative oligosaccharides using the neoglycolipid (NGL) technology; methylation analysis of oligosaccharides to derive glucose linkage information, and incorporation of the newly generated glucan oligosaccharide probes into microarrays encompassing diverse mammalian-type and exogenous sequences for microarray analysis of Dectin-1.
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References
Ariizumi, K., Shen, G. L., Shikano, S., Xu, S., Ritter, R., III, Kumamoto, T., Edelbaum, D., Morita, A., Bergstresser, P. R., and Takashima, A. (2000) Identification of a novel, dendritic cell-associated molecule, dectin-1, by subtractive cDNA cloning, J. Biol. Chem. 275, 20157–20167.
Brown, G. D. and Gordon, S. (2001) Immune recognition. A new receptor for beta-glucans, Nature 413, 36–37.
Brown, G. D., Taylor, P. R., Reid, D. M., Willment, J. A., Williams, D. L., Martinez-Pomares, L., Wong, S. Y., and Gordon, S. (2002) Dectin-1 is a major beta-glucan receptor on macrophages, J. Exp. Med. 196, 407–412.
Brown, G. D., Herre, J., Williams, D. L., Willment, J. A., Marshall, A. S., and Gordon, S. (2003) Dectin-1 mediates the biological effects of beta-glucans, J. Exp. Med. 197, 1119–1124.
Weis, W. I., Taylor, M. E., and Drickamer, K. (1998) The C-type lectin superfamily in the immune system, Immunol. Rev. 163, 19–34.
Willment, J. A., Marshall, A. S., Reid, D. M., Williams, D. L., Wong, S. Y., Gordon, S., and Brown, G. D. (2005) The human beta-glucan receptor is widely expressed and functionally equivalent to murine Dectin-1 on primary cells, Eur. J. Immunol. 35, 1539–1547.
Palma, A. S., Feizi, T., Zhang, Y., Stoll, M. S., Lawson, A. M., Diaz-Rodríguez, E., Campanero-Rhodes, A. S., Costa, J., Brown, G. D., and Chai, W. (2006) Ligands for the beta-glucan receptor, Dectin-1, assigned using ‘designer’ microarrays of oligosaccharide probes (neoglycolipids) generated from glucan polysaccharides, J Biol. Chem. 281, 5771–5779.
Fukui, S., Feizi, T., Galustian, C., Lawson, A. M., and Chai, W. (2002) Oligosaccharide microarrays for high-throughput detection and specificity assignments of carbohydrate-protein interactions, Nat. Biotechnol. 20, 1011–1017.
Feizi, T. and Chai, W. (2004) Oligosaccharide microarrays to decipher the glyco code, Nat. Rev. Mol. Cell Biol. 5, 582–588.
Schallus, T., Jaeckh, C., Feher, K., Palma, A. S., Liu, Y., Simpson, J. C., Mackeen, M., Stier, G., Gibson, T. J., Feizi, T., Pieler, T., and Muhle-Goll, C. (2008) Malectin - A Novel Carbohydrate-binding Protein of the Endoplasmic Reticulum and a Candidate Player in the Early Steps of Protein N-glycosylation, Mol. Biol. Cell 19, 3404–3414.
Liu, Y., Palma, A. S., and Feizi, T. (2009) Carbohydrate microarrays: key developments in glycobiology, Biol. Chem. 390, 647–656.
Jamas, S., Easson, D. D. J., Ostroff, G. R., and Onderdonk, A. B. (1991) Glucans a novel class of macrophage activating immunomodulators, ACS Symp Ser 469, 44–51.
Black, W. A. P., Cornhill, W. J., Dewar, E. J., and Woodward, F. N. (1951) Manufacture of algal chemicals. III. Laboratory-scale isolation of laminarin from brown marine algae, J Appl. Chem. 501–517.
de la, C. J., Pintor-Toro, J. A., Benitez, T., and Llobell, A. (1995) Purification and characterization of an endo-beta-1,6-glucanase from Trichoderma harzianum that is related to its mycoparasitism, J. Bacteriol. 177, 1864–1871.
Chanda, N. B., Hirst, E. L., and Manners, D. J. (1957) A comparison of isoLichenan and Lichenan from Icelandic Moss (Cetrarai islandica), J. Chem Soc. 1951–1958.
McCleary, B. V. and Matheson, N. K. (1987) Enzymic analysis of polysaccharide structure, Advances in Carbohdydrate Chemistry and Biochemistry 44, 147–276.
de Belder, A. N. (1993) Dextran, in Industrial Gums (Whistler, R. L. and BeMiller, J., Eds.) 2nd ed., pp 399–426, Academic Press, New York.
Haworth, W. N., Hirst, E. L., and isherwood, F. A. (1937) Polysaccharides. Part XXIV. Yeast mannan, J Chem Soc 748–790.
Chai, W., Hounsell, E. F., Cashmore, G. C., Rosankiewicz, J. R., Bauer, C. J., Feeney, J., Feizi, T., and Lawson, A. M. (1992) Neutral oligosaccharides of bovine submaxillary mucin. A combined mass spectrometry and 1H-NMR study, Eur. J Biochem. 203, 257–268.
Ciucanu, I. and Kerek, F. (1984) A simple and rapid method for the permethylation of carbohydrates., Carbohydr. Res. 131, 209–217.
Hounsell, E. F., Lawson, A. M., Stoll, M. S., Kane, D. P., Cashmore, G. C., Carruthers, R. A., Feeney, J., and Feizi, T. (1989) Characterisation by mass spectrometry and 500-MHz proton nuclear magnetic resonance spectroscopy of penta- and hexasaccharide chains of human foetal gastrointestinal mucins (meconium glycoproteins), Eur. J. Biochem. 186, 597–610.
Graham, L. M., Tsoni, S. V., Willment, J. A., Williams, D. L., Taylor, P. R., Gordon, S., Dennehy, K., and Brown, G. D. (2006) Soluble Dectin-1 as a tool to detect beta-glucans, J. Immunol. Methods 314, 164–169.
Stoll, M. S., Feizi, T., Loveless, R. W., Chai, W., Lawson, A. M., and Yuen, C. T. (2000) Fluorescent neoglycolipids. Improved probes for oligosaccharide ligand discovery, Eur. J. Biochem. 267, 1795–1804.
Chai, W., Stoll, M. S., Galustian, C., Lawson, A. M., and Feizi, T. (2003) Neoglycolipid technology - deciphering information content of glycome, Methods Enzymol. 362, 160–195.
Leteux, C., Chai, W., Nagai, K., Herbert, C. G., Lawson, A. M., and and Feizi, T. (2001) 10E4 Antigen of scrapie lesions contains an unusual nonsulfated heparan motif., J. Biol. Chem. 276, 12539–12545.
Her, G. R., Santikarn, S., Reinhold, V. N., and Williams, J. C. (1987) Simplified approach to HPLC precolumn fluorescent labeling of carbohydrates:N-(2-pyridinyl)-glycosaylamines, J Carbohydr Chem 6, 129–139.
Liu, Y., Feizi, T., Campanero-Rhodes, M. A., Childs, R. A., Zhang, Y., Mulloy, B., Evans, P. G., Osborn, H. M., Otto, D., Crocker, P. R., and Chai, W. (2007) Neoglycolipid probes prepared via oxime ligation for microarray analysis of oligosaccharide-protein interactions, Chem. Biol. 14, 847–859.
Liu, Y., Chai, W., Childs, R. A., and Feizi, T. (2006) Preparation of Neoglycolipids with Ring-Closed Cores via Chemoselective Oxime-Ligation for Microarray Analysis of Carbohydrate-Protein Interactions, Methods Enzymol. 415C, 326–340.
Palma, A. S., Liu, Y., Muhle-Goll, C., Butters, T. D., Zhang, Y., Childs, R., Chai, W., and Feizi, T. (2010) Multifaceted approaches including neoglycolipid oligosaccharide microarrays to ligand discovery for malectin, Methods Enzymol. 478, 265–286.
Stoll, M. S. and Feizi, T. (2009) Software tools for storing, processing and displaying carbohydrate microarray data. Proceeding of the Beilstein Symposium on Glyco-Bioinformatics, 4–8 October, 2009, Potsdam, Germany, (Kettner, C., Ed.) pp 123–140, Beilstein Institute for the Advancement of Chemical Sciences, Frankfurt, Germany.
Acknowledgments
We gratefully acknowledge collaborations of our colleagues, past and present, in the Glycosciences Laboratory: Mark Stoll, Alex Lawson, and Colin Herbert; and we acknowledge Gordon Brown for the murine Dectin-1-Fc chimera. The Glycosciences Laboratory acknowledges with gratitude collaborators over the years with whom our microarray probes were studied. For grant support, we acknowledge the UK Medical Research Council, the UK Research Councils Basic Technology Grant (GR/S79268, “Glycoarrays”), Engineering and Physical Research Councils Translational Grant EP/G037604/1, and the NCI Alliance of Glycobiologists for Detection of Cancer and Cancer Risk (U01 CA128416). A.S.P. is a fellow of the Fundação para a Ciência e Tecnologia (SFRH/BPD/26515/2006, Portugal).
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Palma, A.S. et al. (2012). Neoglycolipid-Based “Designer” Oligosaccharide Microarrays to Define β-Glucan Ligands for Dectin-1. In: Chevolot, Y. (eds) Carbohydrate Microarrays. Methods in Molecular Biology, vol 808. Humana Press. https://doi.org/10.1007/978-1-61779-373-8_23
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DOI: https://doi.org/10.1007/978-1-61779-373-8_23
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