Characterizing protein-glycosaminoglycan interactions using solution NMR spectroscopy.
Characterizing protein-glycosaminoglycan interactions using solution NMR spectroscopy.
复制标题
使用溶液核磁共振波谱表征蛋白质-糖胺聚糖相互作用。
DOI：
10.1007/978-1-4939-1714-3_26
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Rajarathnam,Krishna
中科院分区：
文献类型：
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作者：
Joseph,PremRajB;Poluri,KrishnaMohan;Sepuru,KrishnaMohan;Rajarathnam,Krishna
溶液核磁共振（NMR）光谱，特别是化学位移微扰（CSP）滴定实验非常适合表征大分子复合物的结合界面。1H-15 N-HSQC为基础的CSP研究已成为选择的方法，由于其简单，时间短的要求，并不需要高层次的NMR专业知识。然而，由于结合诱导的聚集/沉淀和/或质量差的数据，用于表征蛋白质-糖胺聚糖（GAG）相互作用的CSP研究一直具有挑战性。在本章中，我们将讨论如何优化实验变量，如蛋白质浓度，GAG大小，和NMR仪器的灵敏度可以克服这些障碍，以获得有意义的结构见解蛋白质-GAG相互作用。
Solution nuclear magnetic resonance (NMR) spectroscopy and, in particular, chemical shift perturbation (CSP) titration experiments are ideally suited for characterizing the binding interface of macromolecular complexes.1H-15N-HSQC-based CSP studies have become the method of choice due to their simplicity, short time requirements, and not requiring high-level NMR expertise. Nevertheless, CSP studies for characterizing protein–glycosaminoglycan (GAG) interactions have been challenging due to binding-induced aggregation/precipitation and/or poor quality data. In this chapter, we discuss how optimizing experimental variables such as protein concentration, GAG size, and sensitivity of NMR instrumentation can overcome these roadblocks to obtain meaningful structural insights into protein–GAG interactions.