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

这是一项R24提案，旨在建立现代电子自旋共振(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年来非常成功的 国家生物医学中心：ACERT(由NIH P41拨款资助)，其中新的ESR技术 在生物医学合作者的推动下开发，然后提供给广泛的合作者和服务 项目。在作为R24资源的新配置中，它将专注于用户服务和利用 已经建立的广泛和独特的技术已经开发出来。我们有一口井- 为实现这一目标而制定的管理和运作计划。通过外展工作，我们计划 在国内和国际上显著扩大用户基础。我们将提供广泛的教育 以及针对扩大的用户群的培训能力。这将包括一系列研讨会、在线资源、 内部培训和参观其他实验室。外展将包括在主要期刊上发表的教程论文，虚拟的 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 和时事通讯。我们提供的ESR资源在很多方面都是世界上独一无二的。除了我们的 商用CW和脉冲ESR光谱仪覆盖9到35 GHz的频率范围，我们有广泛的 以及独特的定制光谱仪，覆盖9至240 GHz的频率范围。这些 光谱仪致力于生物医学界使用的各种ESR方法，特别是 包括用于传导脉冲偶极ESR(PDS)的技术，其提供标记蛋白质之间的距离 以及可从其推断蛋白质三级结构的RNA位点。它适用于蛋白质复合体。 它们的自然状态。Acert已经开发出两种方法，显著提高了对低分贝的敏感性 微摩尔蛋白质浓度和非常小的样品，它们极大地提高了吞吐量，使 在PDS中研究的许多样本的处理速度更快。虽然PDS提供蛋白质结构，但 常温下多频ESR和脉冲2D-ESR的其他技术提供了详细信息 蛋白质动力学，对学习结构/功能很重要；这包括研究的独特能力 实时纳秒到微秒的动力学，它将解决蛋白质中的结构域运动等问题。 我们创新的软件能够对复杂的一维和二维光谱进行拟合，以提取复杂结构的细节 蛋白质运动，以及我们最近对信号处理软件的开发，导致了独特的软件包 大大提高了微弱ESR信号的信噪比，同时提供了高保真度和精确度 将PDS实验结果转换为(复数)距离分布。这些软件方法将 还可以提高资源的吞吐量。

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.