Jasnin Marion
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
Methods Mol Biol. 2012;836:161-9. doi: 10.1007/978-1-61779-498-8_11.
Cell surface glycosaminoglycans (GAG), such as heparan sulfate (HS) and heparin, are key multifunctional cell regulators, which are involved in numerous molecular events associated with tumor growth, metastasis, pathogen attachment, and immune response. GAG dynamically bind and regulate the activities of many signaling proteins such as growth factors, chemokines, and cytokines. GAG-binding interactions with proteins rely on the coupling between the geometry, flexibility, and rigidity of the polysaccharide chain. Understanding GAG dynamics at the molecular level can therefore provide fundamental insights into GAG function in a cellular context. Elastic incoherent neutron scattering is a powerful tool for the exploration of fast molecular motions in biological macromolecules. Recently, the technique was used to evaluate HS flexibility and rigidity on different timescales between the picosecond (ps) and the nanosecond (ns). Here, neutron spectroscopy experimental procedures are presented, with emphasis on the practical details necessary to prepare samples, run neutron scattering experiments, and extract the dynamics parameters from the data.
细胞表面糖胺聚糖(GAG),如硫酸乙酰肝素(HS)和肝素,是关键的多功能细胞调节剂,参与许多与肿瘤生长、转移、病原体附着和免疫反应相关的分子事件。GAG动态结合并调节许多信号蛋白的活性,如生长因子、趋化因子和细胞因子。GAG与蛋白质的结合相互作用依赖于多糖链的几何形状、柔韧性和刚性之间的耦合。因此,在分子水平上理解GAG动力学可以为其在细胞环境中的功能提供基本见解。弹性非相干中子散射是探索生物大分子中快速分子运动的有力工具。最近,该技术被用于评估皮秒(ps)到纳秒(ns)不同时间尺度上HS的柔韧性和刚性。本文介绍了中子光谱实验程序,重点阐述了制备样品、进行中子散射实验以及从数据中提取动力学参数所需的实际细节。