Analytical Ultracentrifugation as a Tool for Studying Protein Interactions.
Analytical Ultracentrifugation as a Tool for Studying Protein Interactions.
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DOI：
10.1007/s12551-013-0106-2
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发表时间：
2013-06-01
影响因子：
--
通讯作者：
Schuck, Peter
中科院分区：
文献类型：
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作者：
Schuck, Peter
在过去的二十年里，由仪器、理论和计算方法驱动的分析超速离心法领域取得了重大进展。本文将重点介绍沉积平衡(SE)和沉积速度(SV)分析的主要进展。对于SE，这包括分析具有很强热力学非理想性的高浓度示踪剂沉积平衡，以及对于理想相互作用的系统，向具有隐含质量守恒的全球多信号和多速度SE分析发展异质相互作用的分析策略。对于SV，这包括开发和应用Lamm方程的数值解、实现直接边界拟合的噪声分解技术、扩散反卷积沉降系数分布以及多信号沉降系数分布。最近，有效粒子理论揭示了SV中相互作用组分的快速交换系统共迁移的简单物理规律。这为非均质相互作用系统的边界模式的稳健解释开辟了新的可能性。总之，这些SE和SV技术带来了新的方法来研究整个体系中的大分子相互作用，包括稀溶液和高浓度溶液中的亲和力光谱，包括吸引和排斥相互作用，这可以应用于自结合蛋白质的单组分溶液以及多组分溶液中多蛋白质复合体的研究。
The last two decades have led to significant progress in the field of analytical ultracentrifugation driven by instrumental, theoretical, and computational methods. This review will highlight key developments in sedimentation equilibrium (SE) and sedimentation velocity (SV) analysis. For SE, this includes the analysis of tracer sedimentation equilibrium at high concentrations with strong thermodynamic non-ideality, and for ideally interacting systems the development of strategies for the analysis of heterogeneous interactions towards global multi-signal and multi-speed SE analysis with implicit mass conservation. For SV, this includes the development and applications of numerical solutions of the Lamm equation, noise decomposition techniques enabling direct boundary fitting, diffusion deconvoluted sedimentation coefficient distributions, and multi-signal sedimentation coefficient distributions. Recently, effective particle theory has uncovered simple physical rules for the co-migration of rapidly exchanging systems of interacting components in SV. This has opened new possibilities for the robust interpretation of the boundary patterns of heterogeneous interacting systems. Together, these SE and SV techniques have led to new approaches to study macromolecular interactions across the entire the spectrum of affinities, including both attractive and repulsive interactions, in both dilute and highly concentrated solutions, which can be applied to single-component solutions of self-associating proteins as well as the study of multi-protein complex formation in multi-component solutions.