Array-Based Functional Screening of Heparin GlycansReport as inadecuate




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1 AMU - Aix Marseille Université 2 CRCM - Centre de Recherche en Cancérologie de Marseille 3 Institut Paoli-Calmettes 4 University of Liverpool Liverpool 5 LJMU - Liverpool John Moores University

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.





Author: Tania Puvirajesinghe - Yassir Ahmed - Andrew Powell - David Fernig - Scott Guimond - Jeremy Turnbull -

Source: https://hal.archives-ouvertes.fr/



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