Multivalent binding of concanavalin A on variable-density mannoside microarrays

Faraday Discussions
Daniel J VallesAdam B Braunschweig

Abstract

Interactions between cell surface glycans and glycan binding proteins (GBPs) have a central role in the immune response, pathogen-host recognition, cell-cell communication, and a myriad other biological processes. Because of the weak association between GBPs and glycans in solution, multivalent and cooperative interactions in the dense glycocalyx have an outsized role in directing binding affinity and selectivity. However, a major challenge in glycobiology is that few experimental approaches exist for examining and understanding quantitatively how glycan density affects avidity with GBPs, and there is a need for new tools that can fabricate glycan arrays with the ability to vary their density controllably and systematically in each feature. Here, we use thiol-ene reactions to fabricate glycan arrays using a recently developed photochemical printer that leverages a digital micromirror device and microfluidics to create multiplexed patterns of immobilized mannosides, where the density of mannosides in each feature was varied by dilution with an inert spacer allyl alcohol. The association between these immobilized glycans and FITC-labeled concanavalin A (ConA) - a tetrameric GBP that binds to mannosides multivalently - was measure...Continue Reading

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Citations

Apr 22, 2020·Chemistry : a European Journal·Marco MendeFelix F Loeffler
Jul 13, 2019·Faraday Discussions·Marco MendePeter H Seeberger
Sep 26, 2020·Langmuir : the ACS Journal of Surfaces and Colloids·Dimitri WilmsStephan Schmidt
Feb 13, 2020·ACS Applied Materials & Interfaces·Drago StickerJörg P Kutter

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Methods Mentioned

BETA
isothermal titration calorimetry
chip
irradiating
fluorescence microscopy

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ImageJ
Adobe Illustrator

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