Advanced glycosaminoglycan sequencing

蛋白质和细胞表面配体之间的相互作用支撑着许多生物过程，如细胞生长、稳态、凋亡和病原体入侵宿主细胞的能力。一个重要的细胞表面配体家族是几乎在所有哺乳动物细胞表面发现的糖胺聚糖（GAGs）的碳水化合物家族。分析这些gag的线性块状排列序列是一个重大的技术挑战，因为只有极少量的纯材料（容易）可用，化学性质差，检测设备不灵敏。本研究旨在开发和测试一种新的方法来测序这些GAG多糖利用最近的基本改进由申请人。这些发展包括利用分子的另一端，而不是传统使用的（非还原端），具有大大改进的标记机制和先进的检测系统，用于分子的传统端（还原端）。这种方法将提供一种强大而敏感的测序技术，能够利用GAG糖的两端进行表征。从GAG测序中获得的信息将使研究人员能够研究以前难以捉摸的结构和过程，这些结构和过程具有生物学和医学意义。

The interaction between proteins and cell-surface ligands underpin many biological processes such as cell growth, homeostasis, apoptosis and the ability of pathogens to invade host cells. One important family of cell surface ligands are the carbohydrate family of glycosaminoglycans (GAGs) found on the surface of almost all mammalian cells. Analysis of the sequence of the linear block-like arrangement of these GAGs is a significant technical challenge due to only miniscule quantities of pure material being (readily) available, poor chemistries and insensitive detection equipment. This research aims to develop and test a new method for sequencing these GAG polysaccharides utilising recent fundamental improvements made by the applicant. These developments include the exploitation of the opposite end of the molecule than is conventionally used (non-reducing end), with a vastly improved labelling mechanism and an advanced detection system for the conventional end of the molecule (reducing end). This approach will provide a powerful and sensitive sequencing technique capable of employing both ends of the GAG saccharide for characterization. The information gained from GAG sequencing will enable researchers to study previously elusive structures and processes that are biologically and medically significant.