A Glycan Array-Based Assay for the Identification and Characterization of Plant Glycosyltransferases.,Angewandte Chemie International Edition

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A Glycan Array-Based Assay for the Identification and Characterization of Plant Glycosyltransferases.
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2020-05-12 , DOI: 10.1002/anie.202003105
Colin Ruprecht _{1,

2} , Max P Bartetzko _{1,

3} , Deborah Senf _{1,

3} , Anna Lakhina ₄ , Peter J Smith ₄ , Maria J Soto _{4,

5} , Hyunil Oh _{1,

3} , Jeong-Yeh Yang ₄ , Digantkumar Chapla ₄ , Daniel Varon Silva ₁ , Mads H Clausen ₆ , Michael G Hahn ₄ , Kelley W Moremen ₄ , Breeanna R Urbanowicz ₄ , Fabian Pfrengle _{1,

2,

3}

Affiliation

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.

中文翻译：

用于植物糖基转移酶的鉴定和表征的基于聚糖阵列的测定。

种植具有改良细胞壁成分的植物是提高对病原体的抵抗力、增加生物量消化率和调整其他重要特性的有前景的策略。为了改变生物质结构，详细了解细胞壁结构和生物合成是先决条件。我们在这里报告了一种基于聚糖阵列的检测方法，用于植物细胞壁生物合成糖基转移酶（GT）的高通量鉴定和表征。我们证明，不同的异源表达的半乳糖基、岩藻糖基和木糖基转移酶可以将叠氮基功能化的糖核苷酸供体转移到阵列上选定的合成植物细胞壁寡糖，并且转移的单糖可以通过 1,3-与炔基修饰染料的偶极环加成反应。同时筛选数千种假定 GT、核苷酸糖供体和寡糖受体组合的机会将极大地加速植物细胞壁生物合成研究。

更新日期：2020-07-13

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