Turnbull Jeremy E, Miller Rebecca L, Ahmed Yassir, Puvirajesinghe Tania M, Guimond Scott E
Centre for Glycobiology, School of Biological Sciences, University of Liverpool, Liverpool, United Kingdom.
Methods Enzymol. 2010;480:65-85. doi: 10.1016/S0076-6879(10)80004-7.
The heparan sulfate (HS) family of glycosaminoglycans are highly complex and structurally diverse polysaccharides with information encoded within the chains that imparts the ability to bind selectively to a wide range of proteins-the "HS interactome"-and to regulate their biological activities. However, there are two key questions which need to be addressed; first, the extent of structural variation of expressed HS structures-the "heparanome"-in specific biological contexts and second, the degree of functional selectivity exerted by these structures in regulating biological processes. There is a clear need to develop more systematic and high throughput approaches in order to address these questions. Here, we describe a cohort of protocols for profiling different aspects of HS structure and activity, focusing particularly on disaccharide building blocks and larger oligosaccharide domains, the latter representing the functional units of HS chains. A range of other complementary methods in the literature are also discussed. Together these provide a new and more comprehensive toolkit to investigate HS structure and activity in a higher throughput manner in selected biological systems. The implementation of such a glycomics strategy will enable development of a systems biology view of HS structure-function relationships and help to resolve the significant puzzle of the extensive interactome of HS, which remains a key question in the glycobiology field. We anticipate that the next decade will see major advances in our understanding of the complex biology of HS.
硫酸乙酰肝素（HS）糖胺聚糖家族是高度复杂且结构多样的多糖，其链内编码的信息赋予了它们选择性结合多种蛋白质的能力——即“HS相互作用组”，并调节这些蛋白质的生物学活性。然而，有两个关键问题需要解决：第一，在特定生物学背景下，表达的HS结构——“硫酸乙酰肝素组”的结构变异程度；第二，这些结构在调节生物学过程中发挥的功能选择性程度。显然需要开发更系统、高通量的方法来解决这些问题。在此，我们描述了一系列用于分析HS结构和活性不同方面的方案，特别关注二糖构建单元和更大的寡糖结构域，后者代表HS链的功能单元。文中还讨论了文献中的一系列其他互补方法。这些方法共同提供了一个新的、更全面的工具包，以便在选定的生物系统中以更高通量的方式研究HS的结构和活性。实施这样的糖组学策略将有助于形成关于HS结构 - 功能关系的系统生物学观点，并有助于解决HS广泛相互作用组这一重大难题，这仍然是糖生物学领域的一个关键问题。我们预计，未来十年我们对HS复杂生物学的理解将取得重大进展。
