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

项目摘要/摘要 爱荷华大学的研究人员越来越需要分析大分子相互作用和 单分子水平上的构象动力学。十多年来，爱荷华大学卡弗学院 医学、蛋白质和结晶学(P&C)设施一直在为爱荷华大学提供 研究人员接触并帮助操作基本的和最先进的生物物理仪器 以及蛋白质和核酸的结构分析。增加了一种仪器，可以让研究人员 单独、实时、生理学地可视化和研究具有生物学意义的重要分子 这些条件将大大加强爱荷华大学的研究项目。它将提供更多- 需要工具来开发对细胞如何工作和通信、如何 分子机器的组装和运转。虽然爱荷华大学的几个小组已经建立了 实验室的单分子分析系统，这些仪器都不允许同时进行 生物大分子的可视化和操控。目前，也没有单分子 仪器集中在爱荷华大学的非专家用户可用。 这项提议要求为LUMICKS C-Trap(R)仪器提供资金，这是第一个也是唯一一个商业 结合高分辨率的大分子相互作用的单分子分析仪器 光学镊子、荧光显微镜和微流体，因此允许同时 大分子复合体的操纵和可视化。对于单人-它是一个重要的工具- 大分子相互作用、分子组装和功能的分子多参数分析 蛋白质和核酸的机器和构象动力学。 C-Trap(R)将被安置在P&C设施，其人员将负责仪器 维护、用户培训，并将协助调查人员进行实验设计、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- 资助爱荷华大学及其合作者的研究项目，并将成为重要的 为我们在区域机构中的合作者提供资源。将立即从中受益的项目 仪器包括对分子过程的研究，以确保基因组的稳定性、组装和 多组分分子机器、病毒基因组折叠、染色质组织的动力学 作为代表新抗癌、抗神经退行性变、 抗病毒和抗微生物疗法。

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.