Emory Comprehensive Glycomics Center, Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, U.S.A.
Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, U.S.A.
Biochem Soc Trans. 2019 Feb 28;47(1):1-11. doi: 10.1042/BST20170487. Epub 2019 Jan 9.
Glycans in polysaccharides and glycoconjugates of the hydrophilic exterior of all animal cells participate in signal transduction, cellular adhesion, intercellular signaling, and sites for binding of pathogens largely through protein-glycan interactions. Microarrays of defined glycans have been used to study the binding specificities of biologically relevant glycan-binding proteins (GBP), but such arrays are limited by their lack of diversity or relevance to the GBP being investigated. Shotgun glycan microarrays are made up of structurally undefined glycans that were released from natural sources, labeled with bifunctional reagents so that they can be monitored during their purification using multidimensional chromatographic procedures, stored as a tagged glycan library (TGL) and subsequently printed onto microarrays at equal molar concentrations. The shotgun glycan microarray is then interrogated with a biologically relevant GBP and the corresponding glycan ligands can be retrieved from the TGL for detailed structural analysis and further functional analysis. Shotgun glycomics extended the defined glycan microarray to a discovery platform that supports functional glycomic analyses and may provide a useful process for ultimately defining the human glycome.
所有动物细胞亲水性外表面的多糖和糖缀合物中的聚糖参与信号转导、细胞黏附、细胞间信号传递以及病原体结合位点，主要通过蛋白-聚糖相互作用。已使用定义明确的聚糖微阵列来研究生物相关糖结合蛋白 (GBP) 的结合特异性，但此类微阵列受到其缺乏多样性或与正在研究的 GBP 缺乏相关性的限制。 糖组学的“shotgun”聚糖微阵列由从天然来源释放的结构未定义的聚糖组成，并用双功能试剂标记，以便可以在多维色谱过程中对其进行纯化时对其进行监测，以标签聚糖库 (TGL) 的形式储存，并随后以等摩尔浓度打印到微阵列上。然后，用生物相关的 GBP 询问“shotgun”聚糖微阵列，并且可以从 TGL 中检索到相应的糖配体，以进行详细的结构分析和进一步的功能分析。糖组学的“shotgun”方法将定义明确的聚糖微阵列扩展到一个发现平台，该平台支持功能糖组学分析，并且可能为最终定义人类糖组提供有用的方法。
