Shotgun functional glycomics of heparan sulphate saccharides: generating diverse libraries to decode biological selectivity

复合糖(多糖)是一个高度多样化的分子家族，在生物过程中具有广泛的功能，包括细胞识别、黏附、细胞间通信和信号传递。对糖的研究--由特定细胞或组织表达的一整套多糖--是一个新兴领域，旨在了解糖的功能如何支撑人类生物学的复杂性，以及它们在疾病过程中的参与。蛋白质与多糖的选择性相互作用是其生物学功能的关键之一。出现了许多新技术来支持“糖组学”研究的发展--大规模研究多聚糖结构多样性，以及多聚糖与其匹配的蛋白质伙伴的选择性相互作用，这是其功能的基础。然而，这些方法并没有直接揭示功能特性，特别是对于某些复杂类别的多糖，如硫酸乙酰肝素(HS)家族的硫酸多糖，它们负责调节广泛的生物过程，包括生长因子信号、酶活性和细胞黏附。我们现在需要有工具来研究特定的结构如何控制特定的蛋白质来控制生物系统。在这个项目中，我们建议开发一种新的方法来解决这个问题，方法是生成一个具有广泛结构的新型HS糖链的大型文库，并在生物检测中筛选它们，然后对选定的结构进行分析。这将使我们能够第一次评估HS在较高吞吐量下的结构-活性关系。这种‘鸟枪式’文库方法将通过允许特定的糖链结构与特定的生物功能相匹配，为破译HS糖蛋白的功能提供一个强大且通用的新工具。在未来，这一战略可以提供新的信息，可转化为生物技术和药物开发方面的应用。

Complex sugars (glycans) are a highly diverse family of molecules with a broad range of functions in biological processes including cell recognition, adhesion, cell-cell communication and signalling. The study of the glycome - the entire set of glycans expressed by particular cells or tissues - is an emerging field which aims to understand how glycan functions underpin the complexity of human biology, and their involvement in disease processes. The selective interaction of proteins with glycans is one of the keys to their biological functions. A number of new technologies have emerged to support the development of 'glycomics' studies - large-scale studies of glycan structural diversity and the selective interactions of glycans with their matching protein partners which underpins their functions. However, these approaches do not directly reveal functional properties, especially for some complex classes of glycans like the heparan sulphate (HS) family of sulphated glycans which are responsible for regulation of a wide range of biological processes including growth factor signalling, enzyme activity and cell adhesion. We now need to have the tools to ask how specific structures control specific proteins to control biological systems. In this project we propose to develop a new approach to address this question by generating a large library of novel HS glycans with a wide range of structures, and screening them in biological assays, followed by analysis of selected structures. This will permit us to evaluate the structure-activity relationships of HS at higher throughput for the first time. This 'shotgun' library approach will provide a powerful and generic new tool for decoding the function of the HS glycome by allowing specific glycan structures to be matched to specific biological functions. In the future this strategy could provide new information that could be translated into applications in biotechnology and drug development.