Tracking developmentally regulated post-synthetic processing of homogalacturonan and chitin using reciprocal oligosaccharide probes
2014
Mravec, Jozef | Kracun, Stjepan K | Rydahl, Maja G. | Westereng, Bjorge | Miart, Fabien | Clausen, Mads H. | Fangel, Jonatan U | Daugaard, Mathilde | van Cutsem, Pierre | de Fine Licht, Henrik H. de Fine | Höfte, Herman | Malinovsky, Frederikke G | Domozych, David S. | Willats, William G. T. | Department of Plant and Environmental Sciences [Copenhagen] ; Faculty of Science [Copenhagen] ; University of Copenhagen = Københavns Universitet (UCPH)-University of Copenhagen = Københavns Universitet (UCPH) | Dept Chem Biotechnol & Food Sci ; Norwegian University of Life Sciences (NMBU) | Institut Jean-Pierre Bourgin (IJPB) ; Institut National de la Recherche Agronomique (INRA)-AgroParisTech | Ctr Nano Med & Theranost-Dept Chem ; Danmarks Tekniske Universitet = Technical University of Denmark (DTU) | Unite Rech Biol Cellulaire Vegetale ; Université de Namur [Namur] (UNamur) | Dept Bio-Skidmore Microscopy Imaging Ctr ; Skidmore College [Saratoga Springs]
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.
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