Mravec, Jozef4; Kračun, Stjepan K.4; Rydahl, Maja G.4; Westereng, Bjørge4; Miart, Fabien5; Clausen, Mads Hartvig1; Fangel, Jonatan U.4; Daugaard, Mathilde1; Van Cutsem, Pierre6; De Fine Licht, Henrik H.4; Höfte, Herman5; Malinovsky, Frederikke G.4; Domozych, David S.7; Willats, William G. T.4
1 Department of Chemistry, Technical University of Denmark2 Organic Chemistry, Department of Chemistry, Technical University of Denmark3 Center for Nanomedicine and Theranostics, Center, Technical University of Denmark4 University of Copenhagen5 Saclay Plant Sciences6 University of Namur7 Skidmore College
Polysaccharides are major components of extracellular matrices and are often extensively modified post-synthetically to suit local requirements and developmental programmes. However, our current understanding of the spatiotemporal dynamics and functional significance of these modifications is limited by a lack of suitable molecular tools. Here, we report the development of a novel non-immunological approach for producing highly selective reciprocal oligosaccharide-based probes for chitosan (the product of chitin deacetylation) and for demethylesterified homogalacturonan. Specific reciprocal binding is mediated by the unique stereochemical arrangement of oppositely charged amino and carboxy groups. Conjugation of oligosaccharides to fluorophores or gold nanoparticles enables direct and rapid imaging of homogalacturonan and chitosan with unprecedented precision in diverse plant, fungal and animal systems. We demonstrated their potential for providing new biological insights by using them to study homogalacturonan processing during Arabidopsis thaliana root cap development and by analyzing sites of chitosan deposition in fungal cell walls and arthropod exoskeletons.
Development (cambridge), 2014, Vol 141, Issue 24, p. 4841-4850