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A Glycan Array‐Based Assay for the Identification and Characterization of Plant Glycosyltransferases
Authors:Colin Ruprecht  Max P Bartetzko  Deborah Senf  Anna Lakhina  Peter J Smith  Maria J Soto  Hyunil Oh  Jeong‐Yeh Yang  Digantkumar Chapla  Daniel Varon Silva  Mads H Clausen  Michael G Hahn  Kelley W Moremen  Breeanna R Urbanowicz  Fabian Pfrengle
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
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