Tracking developmentally regulated post-synthetic processing of homogalacturonan and chitin using reciprocal oligosaccharide probes.
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.
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Jozef
J
Mravec
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Stjepan K
SK
Kračun
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Maja G
MG
Rydahl
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Aas NO-1432, Norway.
Bjørge
B
Westereng
Institut Jean-Pierre Bourgin, UMR1318 INRA/AgroParisTech, Saclay Plant Sciences, INRA Centre de Versailles, Versailles 78026, Cedex, France.
Fabien
F
Miart
Center for Nano medicine and Theranostics and Department of Chemistry, Technical University of Denmark, Kongens Lyngby DK-2800, Denmark.
Mads H
MH
Clausen
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Jonatan U
JU
Fangel
Center for Nano medicine and Theranostics and Department of Chemistry, Technical University of Denmark, Kongens Lyngby DK-2800, Denmark.
Mathilde
M
Daugaard
Unité de Recherche en Biologie cellulaire végétale, University of Namur, Namur B-5000, Belgium.
Pierre
P
Van Cutsem
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Henrik H
HH
De Fine Licht
Institut Jean-Pierre Bourgin, UMR1318 INRA/AgroParisTech, Saclay Plant Sciences, INRA Centre de Versailles, Versailles 78026, Cedex, France.
Herman
H
Höfte
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark.
Frederikke G
FG
Malinovsky
Department of Biology and Skidmore Microscopy Imaging Center, Skidmore College, Saratoga Springs, NY 12866, USA.
David S
DS
Domozych
Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg 1871, Denmark willats@plen.ku.dk.
William G T
WG
Willats
D017360:Arabidopsis / Q000254:growth & development / Q000378:metabolism; D002473:Cell Wall / Q000648:ultrastructure; D002686:Chitin / Q000302:isolation & purification / Q000378:metabolism*; D058114:Desmidiales / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism*; D053768:Metal Nanoparticles; D046228:Microarray Analysis; D046529:Microscopy, Electron, Transmission; D015335:Molecular Probes / Q000378:metabolism; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D061848:Optical Imaging / Q000379:methods; D010368:Pectins / Q000302:isolation & purification / Q000378:metabolism*; D018518:Plant Root Cap / Q000254:growth & development / Q000378:metabolism
D015335:Molecular Probes; D009844:Oligosaccharides; D002686:Chitin; D010368:Pectins; C003181:polygalacturonic acid
Arabidopsis; Carbohydrate microarrays; Cell wall; Exoskeletons; Fluorescence imaging; Polysaccharides; Root cap