NMR and small-angle scattering-based structural analysis of protein complexes in solution
NMR and small-angle scattering-based structural analysis of protein complexes in solution
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DOI：
10.1016/j.jsb.2010.11.004
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发表时间：
2011-03-01
影响因子：
3
通讯作者：
Sattler, Michael
中科院分区：
文献类型：
--
作者：
Madl, Tobias;Gabel, Frank;Sattler, Michael
多结构域蛋白质复合物的结构分析是当前生物学的一个关键挑战，也是理解基本细胞过程的分子基础的先决条件。溶液技术的使用对于表征这些复合物的域和亚基的四元排列和动力学非常重要。在这方面，解决方案核磁共振是唯一能够进行原子或残基分辨率结构测定和研究多结构域蛋白质及其复合物动态特性的技术。由于大型蛋白质复合物的实验 NMR 数据很少，因此将这些数据与其他解决方案技术的附加信息结合起来是有利的。在此，回顾了将溶液态 NMR 与小角 X 射线和中子散射 (SAXS/SANS) 实验相结合进行大型蛋白质复合物结构分析的实用性和计算方法。结合 NMR 和 SAS 的实验和计算方法的最新进展进行了讨论，并通过文献中的最新示例进行了说明。结合 NMR 和 SAS 数据来研究多结构域蛋白质（即通过柔性接头连接弱相互作用结构域）的互补方面，通过对与九尿苷 (U9) RNA 寡核苷酸结合的人剪接因子 U2AF65 的串联 RNA 识别基序 (RRM) 结构域 (RRM1-RRM2) 的结构分析来说明。 (C) 2010 Elsevier Inc. 保留所有权利。
Structural analysis of multi-domain protein complexes is a key challenge in current biology and a prerequisite for understanding the molecular basis of essential cellular processes. The use of solution techniques is important for characterizing the quaternary arrangements and dynamics of domains and subunits of these complexes. In this respect solution NMR is the only technique that allows atomic- or residue-resolution structure determination and investigation of dynamic properties of multi-domain -proteins and their complexes. As experimental NMR data for large protein complexes are sparse, it is advantageous to combine these data with additional information from other solution techniques. Here, the utility and computational approaches of combining solution state NMR with small-angle X-ray and Neutron scattering (SAXS/SANS) experiments for structural analysis of large protein complexes is reviewed. Recent progress in experimental and computational approaches of combining NMR and SAS are discussed and illustrated with recent examples from the literature. The complementary aspects of combining NMR and SAS data for studying multi-domain proteins, i.e. where weakly interacting domains are connected by flexible linkers, are illustrated with the structural analysis of the tandem RNA recognition motif (RRM) domains (RRM1-RRM2) of the human splicing factor U2AF65 bound to a nine-uridine (U9) RNA oligonucleotide. (C) 2010 Elsevier Inc. All rights reserved.