Array-Based Functional Screening of Heparin Glycans

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• 作者：Tania&#160 ; M. Puvirajesinghe¹ ; ² ; Yassir&#160 ; A. Ahmed¹ ; Andrew&#160 ; K. Powell¹ ; ³ ; David&#160 ; G. Fernig¹ ; Scott&#160 ; E. Guimond¹ ; Jeremy&#160 ; E. Turnbull¹ ; ^{j.turnbull@liverpool.ac.uk}
• 刊名：Chemistry & Biology
• 出版年：2012
• 期刊代码：20_10745521
• 类别：ch
• 出版时间：25 May, 2012
• 卷：19
• 期：5
• 页码：553-558
• 文件大小：579 K

摘要

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Summary

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聽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 鈥済lycobioarray鈥?platform has significant potential as a generic tool for functional glycomics screening.