Array-Based Functional Screening of Heparin Glycans

Tania Puvirajesinghe (1, 2) , Yassir G Ahmed (2) , Andrew G Powell (3, 2) , David G Fernig (2) , Scott E Guimond (2) , Jeremy E Turnbull (2)

1 CRCM - Centre de Recherche en Cancérologie de Marseille
2 University of Liverpool
3 LJMU - Liverpool John Moores University

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.

Domaines: Biologie cellulaire

Tania Puvirajesinghe, Yassir G Ahmed, Andrew G Powell, David G Fernig, Scott E Guimond, et al.. Array-Based Functional Screening of Heparin Glycans. Chemistry and Biology, 2012, 19 (5), pp.553-558. ⟨10.1016/j.chembiol.2012.03.011⟩. ⟨hal-01447680⟩