沉降平衡分析法分析大分子相互作用。
The analysis of macromolecular interactions by sedimentation equilibrium.
机构信息
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0540, USA.
出版信息
Methods. 2011 May;54(1):145-56. doi: 10.1016/j.ymeth.2010.12.005. Epub 2010 Dec 16.
Abstract
The study of macromolecular interactions by sedimentation equilibrium is a highly technical method that requires great care in both the experimental design and data analysis. The complexity of the interacting system that can be analyzed is only limited by the ability to deconvolute the exponential contributions of each of the species to the overall concentration gradient. This is achieved in part through the use of multi-signal data collection and the implementation of soft mass conservation. We illustrate the use of these constraints in SEDPHAT through the study of an A+B+B⇌AB+B⇌ABB system and highlight some of the technical challenges that arise. We show that both the multi-signal analysis and mass conservation result in a precise and robust data analysis and discuss improvements that can be obtained through the inclusion of data from other methods such as sedimentation velocity and isothermal titration calorimetry.
摘要
沉降平衡法研究大分子相互作用是一种高度技术化的方法，无论是在实验设计还是数据分析中都需要非常谨慎。可以分析的相互作用系统的复杂性仅受能够解析每个物种对整体浓度梯度的指数贡献的能力限制。这部分通过使用多信号数据收集和实施软质量守恒来实现。我们通过研究 A+B+B ⇌ AB+B ⇌ ABB 系统来说明这些约束在 SEDPHAT 中的使用，并强调了出现的一些技术挑战。我们表明，多信号分析和质量守恒都导致了精确和稳健的数据分析，并讨论了通过包括来自其他方法（如沉降速度和等温滴定量热法）的数据可以获得的改进。
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