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A Glycan Array-Based Assay for the Identification and Characterization of Plant Glycosyltransferases |
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| Authors: | Dr Colin Ruprecht Dr Max P Bartetzko Dr Deborah Senf Dr Anna Lakhina Dr Peter J Smith Dr Maria J Soto Hyunil Oh Dr Jeong-Yeh Yang Dr Digantkumar Chapla Dr Daniel Varon Silva Prof Dr Mads H Clausen Prof Dr Michael G Hahn Prof Dr Kelley W Moremen Prof Dr Breeanna R Urbanowicz Prof Dr Fabian Pfrengle |
| Institution: | 1. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476 Potsdam, Germany
Present address: Department of Chemistry, University of Natural Resources and Life Sciences Vienna, Muthgasse 18, 1190 Vienna, Austria;2. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476 Potsdam, Germany Institute of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin, Germany;3. Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, Athens, GA, 30602 USA;4. Complex Carbohydrate Research Center, University of Georgia, 315 Riverbend Road, Athens, GA, 30602 USA Present address: US Department of Energy Joint Genome Institute (JGI), Berkeley, CA, 94702 USA;5. Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476 Potsdam, Germany;6. Center for Nanomedicine and Theranostics, Department of Chemistry, Technical University of Denmark, Kemitorvet 207, 2800 Kgs. Lyngby, Denmark |
| Abstract: | Growing plants with modified cell wall compositions is a promising strategy to improve resistance to pathogens, increase biomass digestibility, and tune other important properties. In order to alter biomass architecture, a detailed knowledge of cell wall structure and biosynthesis is a prerequisite. We report here a glycan array-based assay for the high-throughput identification and characterization of plant cell wall biosynthetic glycosyltransferases (GTs). We demonstrate that different heterologously expressed galactosyl-, fucosyl-, and xylosyltransferases can transfer azido-functionalized sugar nucleotide donors to selected synthetic plant cell wall oligosaccharides on the array and that the transferred monosaccharides can be visualized “on chip” by a 1,3-dipolar cycloaddition reaction with an alkynyl-modified dye. The opportunity to simultaneously screen thousands of combinations of putative GTs, nucleotide sugar donors, and oligosaccharide acceptors will dramatically accelerate plant cell wall biosynthesis research. |
| Keywords: | carbohydrates glycan array glycosyltransferases plant cell wall sugar nucleotides |