Array-based functional screening of heparin glycans |
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Authors: | Puvirajesinghe Tania M Ahmed Yassir A Powell Andrew K Fernig David G Guimond Scott E Turnbull Jeremy E |
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Affiliation: | 1. Centre for Glycobiology, Department of Biochemistry and Cell Biology, Institute of Integrative Biology, The University of Liverpool, Liverpool L69 7ZB, UK |
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Abstract: | Array methodologies have become powerful tools for interrogation of glycan-protein interactions but have critically lacked the ability to generate cell response data. Here, we report the development of a slide-based array method exemplified by measurement of activation of fibroblast growth factor signaling by heparin saccharides. Heparan sulfate-deficient Swiss 3T3 cells were overlaid onto an aminosilane-coated slide surface onto which heparin saccharides had been spotted and immobilized. The cells were transiently stimulated with FGF2 and immunofluorescence measured to assess downstream ERK1/2 phosphorylation. Activation of this signaling pathway response was restricted to cells exposed to heparin saccharides competent to activate FGF2 signaling. Differential activation of the overlaid cells by different-sized heparin saccharides was demonstrated by quantitative measurement of fluorescence intensity. This "glycobioarray" platform has significant potential as a generic tool for functional glycomics screening. |
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