Hannover Medical School, Institute for Biophysical Chemistry, Hannover, Germany.
Methods Mol Biol. 2021;2263:397-421. doi: 10.1007/978-1-0716-1197-5_19.
Analytical ultracentrifugation is a powerful tool to characterize interactions of macromolecules in solution. In sedimentation velocity experiments, the sedimentation of interaction partners and complexes can be monitored directly and can be used to characterize interactions quantitatively. As an example, we show how the interaction of the clamp loader subcomplex of DNA polymerase III from E. coli and a template/primer DNA saturated with single-stranded DNA-binding protein can be analyzed by analytical ultracentrifugation with fluorescence detection.
分析超速离心是一种强大的工具,可用于表征溶液中大分子的相互作用。在沉降速度实验中,可以直接监测相互作用伙伴和复合物的沉降,并可用于定量表征相互作用。例如,我们展示了如何通过分析带有荧光检测的超速离心来分析大肠杆菌 DNA 聚合酶 III 的夹钳加载子亚基与单链 DNA 结合蛋白饱和的模板/引物 DNA 之间的相互作用。