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Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research

Authors :
Maja Gro Rydahl
Robert A. Field
Christian Ruzanski
Mads Hartvig Clausen
Barry McCleary
Marie-Christine Ralet
Mathias Christian Franch Andersen
Susan E. Marcus
J. Paul Knox
Henriette L. Pedersen
Laura von Schantz
Vladimír Farkaš
Mats Ohlin
Jonatan U. Fangel
William G.T. Willats
Dept Plant Biol & Biotechnol
University of Copenhagen = Københavns Universitet (KU)
Megazyme International
John Innes Centre
Unité de recherche sur les Biopolymères, Interactions Assemblages (BIA)
Institut National de la Recherche Agronomique (INRA)
Slovak Academy of Sciences (SAS)
Lund University [Lund]
University of Leeds
Technical University of Denmark [Lyngby] (DTU)
Danish Research Council
Scientific Grant Agency (VEGA) (Slovakia) [2/0011/09]
Danish Research Council for Strategic Research
Source :
Journal of Biological Chemistry, Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (47), ⟨10.1074/jbc.M112.396598⟩, Journal of Biological Chemistry 47 (287), . (2012)
Publication Year :
2012
Publisher :
HAL CCSD, 2012.

Abstract

International audience; Microarrays are powerful tools for high throughput analysis, and hundreds or thousands of molecular interactions can be assessed simultaneously using very small amounts of analytes. Nucleotide microarrays are well established in plant research, but carbohydrate microarrays are much less established, and one reason for this is a lack of suitable glycans with which to populate arrays. Polysaccharide microarrays are relatively easy to produce because of the ease of immobilizing large polymers noncovalently onto a variety of microarray surfaces, but they lack analytical resolution because polysaccharides often contain multiple distinct carbohydrate substructures. Microarrays of defined oligosaccharides potentially overcome this problem but are harder to produce because oligosaccharides usually require coupling prior to immobilization. We have assembled a library of well characterized plant oligosaccharides produced either by partial hydrolysis from polysaccharides or by de novo chemical synthesis. Once coupled to protein, these neoglycoconjugates are versatile reagents that can be printed as microarrays onto a variety of slide types and membranes. We show that these microarrays are suitable for the high throughput characterization of the recognition capabilities of monoclonal antibodies, carbohydrate-binding modules, and other oligosaccharide-binding proteins of biological significance and also that they have potential for the characterization of carbohydrate-active enzymes.

Details

Language :
English
ISSN :
00219258 and 1083351X
Database :
OpenAIRE
Journal :
Journal of Biological Chemistry, Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2012, 287 (47), ⟨10.1074/jbc.M112.396598⟩, Journal of Biological Chemistry 47 (287), . (2012)
Accession number :
edsair.doi.dedup.....520605d28cdfa64e337b9846e194e871
Full Text :
https://doi.org/10.1074/jbc.M112.396598⟩