Abstract
Glycans are involved in a variety of physiological and pathological processes through interactions with proteins. Thus, the molecular basis of glycan–protein interactions provides valuable information on understanding biological phenomena and exploiting more effective carbohydrate-based therapeutic agents and diagnostic tools. Carbohydrate microarray technology has become a powerful tool for evaluating glycan-mediated biological events in a high-throughput manner. This technology is mostly applied for rapid analysis of glycans–protein interactions in the field of functional glycomics. In order to expand application areas of glycan microarrays, we have used carbohydrate microarrays for measurement of binding affinities between glycans and proteins and profiling of glycosyltransferase activities. The glycan microarrays used for these studies are constructed by immobilizing maleimide or hydrazide-conjugated glycans on the thiol or hydrazide-derivatized glass slides, respectively. This protocol describes the fabrication of carbohydrate microarrays and their applications to enzymatic reactions and determination of quantitative binding affinities.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bertozzi, C. R., and Kiessling, L. L. (2001) Chemical glycobiology. Science 291, 2357–2364.
Varki, A. (1993) Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3, 97–130.
Roth, J. (2002) Protein N-glycosylation along the secretory pathway: relationship to organelle topography and function, protein quality control, and cell interactions. Chem. Rev. 102, 285–303.
Park, S., Lee, M.-R., and Shin, I. (2008) Chemical tools for functional studies of glycans. Chem. Soc. Rev. 37, 1579–1591.
Smith, A. E., and Helenius, A. (2004) How viruses enter animal cells. Science 304, 237–242.
Fuster, M. M., and Esko, J. D. (2005) The sweet and sour of cancer: glycans as novel therapeutic targets. Nat. Rev. Cancer 5, 526–542.
Lasky, L. A. (1992) Selectins: interpreters of cell-specific carbohydrate information during inflammation. Science 258, 964–969.
Scanlan, C.N., Offer, J., Zitzmann, N., and Dwek, R.A. (2007) Exploiting the defensive sugars of HIV-1 for drug and vaccine design. Nat. Insight 446, 1038–1045.
Lis, H., and Sharon, N. (1972) Lectins: cell-agglutinating and sugar-specific proteins. Science 177, 949–959.
McCoy Jr, J. P., Varani, J., and Goldstein, I. J. (1983) Enzyme-linked lectin assay (ELLA): use of alkaline phosphatase-conjugated Griffonia simplicifolia B4 isolectin for the detection of alpha-D-galactopyranosyl end groups. Anal. Biochem. 130, 437–444.
Duverger, E., Frison, N., Roche, A. C., and Monsingny, M. (2003) Carbohydrate-lectin interactions assessed by surface plasmon resonance. Biocheimie 85, 167–179.
Dam, T. K., and Brewer, C. F. (2002) Thermodynamic studies of lectin-carbohydrate interactions by isothermal titration calorimetry. Chem. Rev. 102, 387–430.
Park, S., and Shin, I. (2002) Fabrication of carbohydrate chips for studying protein-carbohydrate interactions. Angew. Chem. Int. Ed. 41, 3180–3182.
Wang, D., Liu, S., Trummer, B. J., Deng, C., and Wang, A. (2002) Carbohydrate microarrays for the recognition of cross-reactive molecular markers of microbes and host cells. Nat. Biotechnol. 20, 275–281.
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.
Houseman, B. T., and Mrksich, M. (2002) Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. Chem. Biol. 9, 443–454.
Park, S., Lee, M.-R., Pyo, S. J., and Shin, I. (2004) Carbohydrate chips for studying high-throughput carbohydrate-protein interactions. J. Am. Chem. Soc. 126, 4812–4819.
Lee, M.-R., and Shin, I. (2005) Fabrication of chemical microarrays by efficient immobilization of hydrazide-linked substances on epoxide-coated glass surfaces. Angew. Chem. Int. Ed. 44, 2881–2884.
Park, S., and Shin, I. (2007) Carbohydrate microarrays for assaying galactosyltransferase activity. Org. Lett. 9, 1675–1678.
Lee, M.-R., and Shin, I. (2005) Facile Preparation of Carbohydrate Microarrays by Site-Specific, Covalent Immobilization of Unmodified Carbohydrates on Hydrazide-Coated Glass Slides. Org. Lett. 7, 4269–4272.
Blixt, O. et al. (2004) Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc. Natl. Acad. Sci. USA 101, 17033–17038.
de Paz, J. L., Noti, C., and Seeberger, P. H. (2006) Microarrays of synthetic heparin oligosaccharides. J. Am. Chem. Soc. 128, 2766–2767.
Disney, M. D., and Seeberger, P. H. (2004) The use of carbohydrate microarrays to study carbohydrate-cell interactions and to detect pathogens. Chem. Biol. 11, 1701–1707.
Park, S., Lee, M.-R., and Shin, I. (2009) Construction of carbohydrate microarrays by using one-step, direct immobilizations of diverse unmodified glycans on solid surfaces. Bioconjugate Chem. 20, 155–162.
Park, S., Lee, M.-R., and Shin, I. (2007) Fabrication of carbohydrate chips and their use to probe protein-carbohydrate interactions. Nat. Protoc. 2, 2747–2758.
Shin, I., Cho, J. W., and Boo, D. W. (2004) Carbohydrate arrays for functional studies of carbohydrates. Comb. Chem. High Throughput Screening 7, 565–574.
Shin, I., Park, S., and Lee, M.-R. (2005) Carbohydrate microarrays: an advanced technology for functional studies of glycans. Chem. Eur. J. 11, 2894–2901.
Shin, I., Tae, J., and Park, S. (2007) Carbohydrate microarray technology for functional glycomics. Curr. Chem. Biol. 1, 187–199.
Shin, I. (2006) Carbohydrate microarrays for high-throughput analysis of carbohydrate-protein interactions. In Protein-Carbohydrate Interactions in Infectious Diseases. (Ed. Carole A. Bewley) 221–46 (RSC Publishing, Cambridge, UK).
Park, S., Lee, M.-R., and Shin, I. (2008) Carbohydrate microarrays as powerful tools in studies of carbohydrate-mediated biological processes. Chem. Commun. 4389–4399.
Horlacher, T., and Seeberger, P. H. (2008) Carbohydrate arrays as tools for research and diagnostics. Chem. Soc. Rev. 37, 1414–1422.
Laurent, N., Voglmeir, J., and Flitsch, S. L. (2008) Glycoarrays-tools for determining protein-carbohydrate interactions and glycoenzyme specificity. Chem. Commun. 4400–4412.
Liang, P.-H., Wang, S.-K., and Wong, C.-H. (2007) Quantitative analysis of carbohydrate-protein interactions using glycan microarrays: determination of surface and solution dissociation constants. J. Am. Chem. Soc. 128 11177–11184.
Acknowledgments
This work was supported by grants of the National Creative Research Initiative and WCU programs.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Lee, MR., Park, S., Shin, I. (2012). Carbohydrate Microarrays for Enzymatic Reactions and Quantification of Binding Affinities for Glycan–Protein Interactions. In: Chevolot, Y. (eds) Carbohydrate Microarrays. Methods in Molecular Biology, vol 808. Humana Press. https://doi.org/10.1007/978-1-61779-373-8_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-373-8_7
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-372-1
Online ISBN: 978-1-61779-373-8
eBook Packages: Springer Protocols