Schuck Peter
Dynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, U.S.A.
Biophys Rev. 2013 Jun 1;5(2):159-171. doi: 10.1007/s12551-013-0106-2.
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.
在仪器、理论和计算方法的推动下，过去二十年分析超速离心领域取得了重大进展。本综述将重点介绍沉降平衡（SE）和沉降速度（SV）分析的关键进展。对于SE，这包括在高浓度且具有强热力学非理想性的情况下对示踪剂沉降平衡的分析，以及对于理想相互作用体系，开发用于分析异质相互作用的策略，以实现具有隐含质量守恒的全局多信号和多速度SE分析。对于SV，这包括Lamm方程数值解的开发与应用、能够直接拟合边界的噪声分解技术、扩散反褶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中相互作用组分快速交换体系共迁移的简单物理规律。这为稳健解释异质相互作用体系的边界模式开辟了新的可能性。总之，这些SE和SV技术带来了新的方法，可用于研究稀溶液和高浓度溶液中亲和力全谱范围内的大分子相互作用，包括吸引和排斥相互作用，既可以应用于自缔合蛋白的单组分溶液，也可以用于研究多组分溶液中多蛋白复合物的形成。
