Multi-Method Approaches for the Study of Complex Protein Interactions

研究复杂蛋白质相互作用的多种方法

• 批准号: 8933882
• 负责人: PETER SCHUCK
• 金额: $ 19.34万
• 依托单位: NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8933882
• 关键词: Affinity; Binding; Biosensing Techniques; Calorimetry; Cells; Complex; Computer software; Data; Development; Educational workshop; Experimental Designs; Feedback; Fluorescence; Goals; Imagery; Medical center; Methods; Modeling; Molecular; Optical Methods; Property; Research Personnel; Signal Transduction; Surface Plasmon Resonance; Titrations; United States National Institutes of Health; analytical tool; analytical ultracentrifugation; biophysical techniques; data acquisition; flexibility; interest; microcalorimetry; protein complex; sedimentation equilibrium; sedimentation velocity; simulation; tool

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.

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。这包括分析超速离心法(沉淀速度和沉淀平衡)、等温滴定微量热法、光谱方法和光学生物传感。具体地说，我们为所有数据类型实现了可视化工具，以将实验数据显示在结合等温线或实验观察值的空间中，作为浓度的函数。这可以突出数据的信息含量，并有助于审查实验设计。此外，我们还实施了广泛的模拟工具，使研究人员能够探索从某种生物物理技术获得的数据是否有希望有助于表征感兴趣的分子参数。 为了传播这一工具并促进其同事的应用，我们在贝塞斯达的NIH、达拉斯大学西南医学中心和汉诺威的Medizinische Hochschule举办了研讨会。这产生了积极的反馈和软件改进。 在实验上，随着荧光检测沉积速度(FDS-SV)方法的发展，我们大大扩展了GMMA的潜力，用于研究高亲和力相互作用。FDS-SV现在提供了在低NM范围内研究与亲和力相互作用的可能性，补充了等温滴定量热法和表面等离子体共振生物传感。