Multi-Method Approaches for the Study of Complex Protein Interactions

我们继续扩展全球多方法分析（GMMA）软件SEDPHAT，该软件为多方法方法中的多组分相互作用分析提供了各种模型和特定的统计功能。这包括分析超离心（沉降速度和沉降平衡），等温滴定微热法，光谱方法和光学生物传感。具体来说，我们已经为所有数据类型实现了可视化工具，以在绑定等温线空间或实验可观测值中显示实验数据，作为浓度的函数。这样可以突出数据的信息内容，有助于检验实验设计。此外，我们已经实施了广泛的模拟工具，允许研究者探索从某种生物物理技术获取数据是否有希望帮助表征感兴趣的分子参数。

We have continued to expand the global multi-method analysis (GMMA) software SEDPHAT, which offers a variety of models and specific statistical functions for the analysis of multi-component interactions in a multi-method approach. This includes analytical ultracentrifugation (sedimentation velocity and sedimentation equilibrium), isothermal titration microcalorimetry, spectroscopic methods, and optical biosensing. Specifically, we have implemented visualization tools for all data types to show experimental data in a space of binding isotherms, or experimental observables, as a function of concentration. This can highlight the information content of data and help examining experimental design. Further, we have implemented extensive simulation tools that allow the investigator to explore whether the acquisition of data from a certain biophysical technique would be promising to aid in the characterization of the molecular parameter of interest. In order to disseminate this tool and facilitate its application by colleagues, we have held workshops at NIH in Bethesda, at the Dallas UT Southwestern Medical Center, and at the Medizinische Hochschule Hannover. This has generated positive feedback and software improvements. Experimentally, we have significantly extended the potential for GMMA with the development of fluorescence-detected sedimentation velocity (FDS-SV) method for the study of of high-affinity interactions. FDS-SV offers now a possibility to study of interactions with affinities in the low nM range, complementary to isothermal titration calorimetry and surface plasmon resonance biosensing.