Correia John J, Stafford Walter F
Department of Biochemistry, University of Mississippi Medical Center, Jackson, Mississippi, USA.
Department of Systems Biology, Boston Biomedical Research Institute and Harvard Medical School, Boston, Massachusetts, USA.
Methods Enzymol. 2015;562:49-80. doi: 10.1016/bs.mie.2015.06.042. Epub 2015 Aug 13.
Here we give an overview of the history of sedimentation velocity analysis focusing on seminal and fundamental contributions that derived from early ultracentrifugation studies. We introduce the concepts of nonequilibrium thermodynamics and outline the derivation of the Svedberg and the Lamm equations and the requirements for including both hydrodynamic and thermodynamic nonideality. We introduce the phenomenological equations for coupled flows as developed from the principles of nonequilibrium or irreversible thermodynamics and derive a form of the Lamm equation that incorporates cross-diffusion coefficients and coupled gradient terms. We give an historical overview of solutions to the Lamm equation including Fujita-MacCosham solutions and Claverie finite-element numerical solutions and discuss the software that have implemented these solutions. We discuss the three major optical systems (absorbance, interference, and fluorescence) and recently developed multiwavelength systems. We also suggest a number of experimental practices and guidelines for optimizing the determination of s and D and discuss the appropriate centerpiece components and their utility. This chapter complements other recent reviews submitted by the authors (Correia, Lyons, Sherwood, & Stafford, 2015; Stafford, 2015) and should be considered an effort to revive the importance of irreversible thermodynamics in the understanding and analysis of sedimentation velocity ultracentrifugation data.
在此，我们概述沉降速度分析的历史，重点关注早期超速离心研究中的开创性和基础性贡献。我们介绍非平衡热力学的概念，概述斯维德贝格方程和拉姆方程的推导，以及纳入流体动力学和热力学非理想性的要求。我们介绍从非平衡或不可逆热力学原理发展而来的耦合流现象学方程，并推导包含交叉扩散系数和耦合梯度项的拉姆方程形式。我们对拉姆方程的求解方法进行历史概述，包括藤田 - 麦克科沙姆解法和克拉维里有限元数值解法，并讨论实现这些解法的软件。我们讨论三种主要的光学系统（吸光度、干涉和荧光）以及最近开发的多波长系统。我们还提出一些优化沉降系数(s)和扩散系数(D)测定的实验方法和指南，并讨论合适的核心组件及其用途。本章补充了作者最近提交的其他综述（科雷亚、莱昂斯、舍伍德和斯塔福德，2015年；斯塔福德，2015年），应被视为一次重振不可逆热力学在理解和分析沉降速度超速离心数据方面重要性的努力。
