Abstract
Glycosaminoglycans (GAGs) perform numerous vital functions within the body. As major components of the extracellular matrix, these polysaccharides participate in a diverse array of cell-signaling events. We have developed a simple microarray assay for the evaluation of protein binding to various GAG subclasses. In a single experiment, the binding to all members of the GAG family can be rapidly determined, giving insight into the relative specificity of the interactions and the importance of specific sulfation motifs. The arrays are facile to prepare from commercially available materials.
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References
Laurent, T. C., Laurent, U. B., and Fraser, J. R. (1996) The structure and function of hyaluronan: an overview, Immunol. Cell Biol. 74, A1–A7.
Iozzo, R. V., and Antonio, J. D. S. (2001) Heparan sulfate proteoglycans: heavy hitters in the angiogenesis arena, J. Clin. Invest. 108, 349–355.
Holt, C. E., and Dickson, B. J. (2005) Sugar codes for axons?, Neuron 46, 169–172.
Thammawat, S., Sadlon, T. A., Hallsworth, P. G., and Gordon, D. L. (2008) Role of cellular glycosaminoglycans and charged regions of viral G protein in human metapneumovirus infection, J. Virol. 82, 11767–11774.
Galtrey, C. M., and Fawcett, J. W. (2007) The role of chondroitin sulfate proteoglycans in regeneration and plasticity in the central nervous system, Brain Res. Rev. 54, 1–18.
Sugahara, K., Mikami, T., Uyama, T., Mizuguchi, S., Nomura, K., and Kitagawa, H. (2003) Recent advances in the structural biology of chondroitin sulfate and dermatin sulfate, Curr. Opin. Struct. Biol. 13, 612–620.
Lui, D., Shriver, Z., Qi, Y., Venkataraman, G., and Sasisekharan, R. (2002) Dynamic regulation of tumor growth and metasis by heparan sulfate glycosaminoglycans, Semin. Thromb. Hemost. 28, 67–78.
TĂmár, J., Lapis, K., Dudás, J., SebestyĂ©n, A., Kopper, L., and Kovalszky, I. (2002) ProteoÂglycans and tumor progression: Janus-faced molecules with contradictory functions in cancer, Semin. Cancer Biol. 12, 173–186.
Sanderson, R. D. (2001) Heparan sulfate proteoglycans in invasion and metastasis, Semin. Cell Dev. Biol. 12, 89–98.
Casu, B., Guerrini, M., and Torri, G. (2004) Structural and conformational aspects of the anticoagulant and anti-thrombotic activity of heparin and dermatan sulfate, Curr. Pharm. Des. 10, 939–949.
Fareed, J., Hoppensteadt, D. A., and Bick, R. L. (2000) An update on heparins at the beginning of the new millennium, Semin. Thromb. Hemost. 26, 5–18.
Capila, I., and Linhardt, R. J. (2002) Heparin-protein interactions, Angew. Chem. Int. Ed. 41, 391–412.
Whitelock, J. M., and Iozzo, R. V. (2005) Heparan sulfate: a complex polymer charged with biological activity, Chem. Rev. 105, 2745–2764.
Rubin, J. B., Choi, Y., and Segal, R. A. (2002) Cerebellar proteoglycans regulate sonic hedgehog responses during development, Development 129, 2223–2232.
Taylor, K. R., and Gallo, R. L. (2006) Glycosaminoglycans and their proteoglycans: host-associated molecular patterns for initiation and modulation of inflammation, FASEB J. 20, 9–22.
Sugahara, K., and Mikami, T. (2007) Chondroitin/dermatan sulfate in the central nervous system, Curr. Opin. Struct. Biol. 17, 536–545.
de Paz, J. L., Noti, C., and Seeberger, P. H. (2006) Microarrays of synthetic heparin oligosaccharides, J. Am. Chem. Soc. 128, 2766–2767.
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.
Gama, C. I., Tully, S. E., Sotogaku, N., Clark, P. M., Rawatand, M., Vaidehi, N., Goddard, W. A., Nishi, A., and Hsieh-Wilson, L. C. (2006) Sulfation patterns of glycosaminoglycans encode molecular recognition and activity, Nat. Chem. Biol. 2, 467–473.
Shipp, E. L., and Hsieh-Wilson, L. C. (2007) Profiling the sulfation specificities of glycosaminoglycan interactions with growth factors and chemotactic proteins using microarrays, Chem. Biol. 14, 195–208.
Tully, S. E., Rawat, M., and Hsieh-Wilson, L. C. (2006) Discovery of a TNF-alpha antagonist using chondroitin sulfate microarrays, J. Am. Chem. Soc. 128, 7740–7741.
Acknowledgments
We thank Dr. Igor Antoshechkin, Director of the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology, and Dr. Jose Luis Riechmann for assistance with printing the microarrays and Joshua M. Brown, Russell E. Roberson, and Dr. Igor Antoshechkin for critically reading the manuscript. This work was supported by the National Institutes of Health (R01 GM093627).
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Rogers, C.J., Hsieh-Wilson, L.C. (2012). Microarray Method for the Rapid Detection of Glycosaminoglycan–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_22
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DOI: https://doi.org/10.1007/978-1-61779-373-8_22
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