National Biomedical Resource for Electron-Spin Resonance Spectroscopy (ACERT)

这是一个R24建议，建立一个现代电子自旋共振（ESR）的区域和国家资源 供生物医学界使用的技术。这一资源将建立在20年的一个非常成功的 国家生物医学中心：ACERT（由NIH P41资助），其中新的ESR技术， 由生物医学合作者驱动开发的，然后提供给广泛的合作者和服务 项目在其作为R24资源的新配置中，它将专注于用户服务和协作， 已经建立的广泛和独特的技术已经开发。我们有一个很好的- 为实现这一目标制定了行政和业务计划。通过外联工作，我们计划 在国内和国际上大大扩大用户基础。我们将提供广泛的教育 以及为扩大的用户群提供培训能力。这将包括一系列的讲习班，在线资源， 内部培训和参观其他实验室。外联活动将包括在主要期刊上发表辅导论文， 与国际ESR协会的会议，YouTube上的视频讲座，以及领先期刊上的广告 和报纸等多个形式分享我们提供的ESR资源在许多方面都是世界上独一无二的。除了我们 我们拥有广泛的商用连续波和脉冲ESR光谱仪，频率范围为9至35 GHz， 和独特的定制家庭内置频谱仪，覆盖9至240 GHz的频率范围。这些 光谱仪专用于生物医学界使用的全方位ESR方法，特别是 包括用于进行脉冲偶极ESR（PDS）的技术，其提供标记的蛋白质之间的距离 以及可以推断蛋白质三级结构的RNA位点。它适用于蛋白质复合物， 他们的自然状态。ACERT已经开发出了既能显著提高对低亚微米的敏感性， 微摩尔蛋白浓度和非常小的样品，它们大大提高了通量， PDS中研究的许多样品的处理速度更快。虽然PDS提供蛋白质结构， 在环境温度下的多频ESR和脉冲2D-ESR的附加技术提供了 蛋白质动力学，在学习结构/功能方面很重要;这包括研究蛋白质动力学的独特能力。 真实的时间纳秒到微秒动力学，这将解决蛋白质中的域运动等问题。 我们的创新软件能够拟合复杂的1-D和2-D光谱，以提取复杂的细节。 蛋白质运动，我们最近开发的信号处理软件已经导致了独特的软件包， 大大增加了弱ESR信号的信噪比，同时提供了很大的保真度，以及准确的 将PDS实验的结果转换为（复杂的）距离分布。这些软件方法将 还增强了资源的吞吐量。

This is an R24 proposal to establish a Regional and National Resource in modern Electron-Spin Resonance (ESR) Technology for use by the biomedical community. This resource will build upon 20 years of a very successful National Biomedical Center: ACERT (funded by an NIH P41 grant), in which the new ESR technologies that were developed, driven by biomedical collaborators, were then provided to extensive collaborators and service projects. In its new configuration as an R24 resource, it will focus on user services and collaborations utilizing the already established extensive and unique technologies that have already been developed. We have a well- established administration and operations scheme for this objective. Through outreach efforts we plan to significantly expand the user base both nationally and internationally. We will be providing extensive educational and training capabilities for the expanded user base. This will include a series of workshops, on-line resources, in house training and visits to other labs. The outreach will include tutorial papers in key journals, virtual meetings with the International ESR Society, video lectures on YouTube, and advertisements in leading Journals and Newsletters. The ESR resources we provide are in many ways unique in the world. In addition to our commercial spectrometers for cw and pulse ESR covering the frequency range of 9 to 35 GHz, we have extensive and unique custom-home built spectrometers covering the range of frequencies 9 to 240 GHz. These spectrometers are dedicated to a full range of ESR methods utilized by the biomedical community, especially including techniques for conducting pulse dipolar ESR (PDS), which provides distances between labeled protein and RNA sites from which protein tertiary structure may be inferred. It is applicable to protein complexes in their natural states. ACERT has developed methods that both significantly increase the sensitivity to low sub- micromolar protein concentrations and very small samples, and they greatly increase the throughput, enabling a greater rate of processing of the many samples studied in PDS. While PDS supplies protein structures, the additional techniques of multi-frequency ESR and pulsed 2D-ESR at ambient temperatures supply details of protein dynamics, important in learning about structure/function; this includes the unique ability to study in real time nanosecond to microsecond dynamics, which will address such issues as domain motions in proteins. Our innovative software enables the fitting of the complex 1- and 2-D spectra to extract the details of the complex protein motions, and our recent developments of signal processing software have led to unique packages for greatly increasing the signal-to-noise of weak ESR signals while providing great fidelity, as well as accurately transforming the results of PDS experiments into (complex) distance distributions. These software methods will also enhance the throughput of the Resource.