Lumick's C-Trap instrument for single-molecule analysis of macromolecular dynamics

项目概要/摘要 爱荷华州大学的研究人员越来越需要分析大分子相互作用， 单分子水平的构象动力学。十多年来，爱荷华州大学卡弗学院 医学蛋白质和晶体学（P & C）设施一直提供爱荷华州大学 研究人员获得并帮助操作基本和最先进的生物物理仪器， 以及蛋白质和核酸的结构分析。增加一种仪器，使研究人员 在真实的时间内，在生理条件下， 条件将大大提高爱荷华州大学的研究项目。它将提供一个非常- 需要一个工具来发展细胞如何工作和交流的机械理解， 分子机器组装和运作。虽然爱荷华州大学的几个小组已经建立了 单分子分析系统，这些仪器都不允许同时 可视化和操纵生物大分子。目前，也没有单分子 仪器集中提供给非专家用户在爱荷华州的大学。 该提案要求为LUMICKS C-Trap（R）仪器提供资金，这是第一个也是唯一一个商业化的仪器。 结合高分辨率的用于大分子相互作用的单分子分析的仪器 光学镊子，荧光显微镜和微流体，因此允许同时 操作和可视化的大分子复合物。它是一个重要的工具，为单一的- 分子间相互作用、组装和功能的分子多参数分析 机器和蛋白质和核酸的构象动力学。 C-Trap（R）将存放在P & C机构，其人员将负责仪器 维护，用户培训，并将协助研究人员与实验设计，仪器 操作和数据分析。C-Trap（R）仪器将对许多NIH产生直接影响- 资助的研究项目在爱荷华州和合作者的大学，也将作为一个重要的 为我们在区域机构的合作者提供资源。将立即从中受益的项目 这些工具包括对确保基因组稳定性、组装和 多组分分子机器的动力学，病毒基因组折叠，染色质组织，以及 作为代表用于开发新的抗癌，抗神经变性， 抗病毒和抗菌治疗。

PROJECT SUMMARY/ABSTRACT Investigators at the University of Iowa have a growing need to analyze macromolecular interactions and conformational dynamics at a single-molecule level. For over a decade, University of Iowa Carver College of Medicine Protein and Crystallography (P&C) Facility has been providing the University of Iowa investigators access to and help in operating of basic and state-of-the-art instrumentation for biophysical and structural analysis of proteins and nucleic acids. Addition of an instrument that will allow researchers to visualize and study biologically important molecules individually, in real time, and under physiological conditions will significantly enhance research programs at the University of Iowa. It will provide a much- needed tool for developing mechanistic understanding of how cells work and communicate, how molecular machines assemble and function. While several groups at the University of Iowa have built systems for single-molecule analysis in their labs, none of these instruments allows for simultaneous visualization and manipulation of biological macromolecules. Currently, there is also no single-molecule instruments centrally available to non-expert users at the University of Iowa. This proposal requests funds for a LUMICKS C-Trap(R) instrument, the first and only commercial instrument for single-molecule analysis of macromolecular interactions that combines high resolution optical tweezers, fluorescence microscopy and microfluidics, and therefore allows simultaneous manipulation and visualization of the macromolecular complexes. It is an important tool for the single- molecule multiparameter analysis of macromolecular interactions, assembly and function of molecular machines and conformational dynamics of proteins and nucleic acids. The C-Trap(R) will be housed at the P&C Facility whose personnel will be responsible for the instrument maintenance, user training, and will assist the investigators with experimental design, instrument operation and data analysis. The C-Trap(R) instrument will have an immediate impact on numerous NIH- funded research projects at the University of Iowa and collaborators, and will also serve as an important resource for our collaborators in regional institutions. The projects that will immediately benefit from this instrument include studies of the molecular processes that ensure genome stability, assembly and dynamics of multicomponent molecular machines, viral genome folding, chromatin organization, as well as enzymes that represent attractive targets for development of new anticancer, anti-neurodegeneration, anti-viral and antimicrobial therapies.