Dynamics of Protein Assemblies by Analytical Ultracentrifugation

我们进一步测试和开发了我们的新工具，使用质量守恒约束作为贝叶斯正则化的形式在多信号沉积速度。 我们已经证实，通过计算机模拟和应用实验数据集，它可以显着地帮助成分的光谱歧视。 我们已经在软件SEDPHAT中完成了这种方法的复杂实现。 为了研究具有相对较宽粒度分布的多糖和其他大分子，我们已经开始与Stephen Harding博士合作，开发基于M-star分析原理和直接全局拟合的新沉降平衡方法，积分方程代表粒度分布。 人们越来越认识到，超弱相互作用可能在高浓度的细胞环境中发挥重要作用。 因此，我们开始了一项利用流体动力学相互作用的测量报告弱大分子相互作用的研究。 我们已经确定了实验模型系统，并正在研究水力和热力学非理想性的理论框架。 为了总结和传播我们和其他人在分析超离心领域开发的新工具，我们为Current Protocols in Protein Science撰写了一篇全面的评论，并举办了两次研讨会来教授先进的分析超离心方法。

We have further tested and developed our new tool of using mass conservation constraints as a form of Bayesian regularization in multi-signal sedimentation velocity. We have confirmed by computer simulations and application to experimental data sets that it can significantly aid in the spectral discrimination of components. We have finished the complex implementation of this approach in the software SEDPHAT. Towards the study of polysaccharides and other macromolecules with relatively broad particle size distribution, we have embarked on a collaboration with Dr. Stephen Harding for the development of new sedimentation equilibrium approaches based on the principles of M-star analysis and on the direct global fitting with integral equations representing size distributions. It is increasingly recognized that ultra-weak interactions may play an important role in the high concentrations of the cellular milieu. Therefore, we initiated a study of exploiting the measurement of hydrodynamic interactions to report on weak macromolecular interactions. We have identified experimental model systems, and are studying the theoretical framework of hydroydamic and thermodynamic non-ideality. In order to summarize and disseminate the new tools developed by us and others in the field of analytical ultracentrifugation, we wrote a comprehensive review for Current Protocols in Protein Science, and conducted two workshops to teach advanced analytical ultracentrifugation methodology.