LABELING HEPATITIS B CORE PARTICLES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 蛋白质和多蛋白质组装体的三维结构的知识是 对理解基本生物过程和设计 更有效的药物。对于许多这样的物种，它们不能结晶， 对于NMR分析太大，电子显微镜（EM）是唯一可以提供 有用的结构信息。电子致密分子的附着， 以高效率散射电子的重金属元素簇，通常是 这是EM数据收集和分析所必需的。高分辨率EM的从业者通常具有 只有两种电子密度高的材料和一套有限的连接方法 可从商业供应商获得。在许多情况下，这些资源失败，目标必须 要么被抛弃，要么被费力地重新设计。我们建议创造新的电子密度 化合物，并开发方法，以快速确定最佳的方式将它们连接到目标 proteins.目标是为EM社区提供一套扩展的材料和技术 用于蛋白质标记，以便更大比例的靶标可以在结构上表征。 这项工作将包括等量的化学合成、化学分析和测量 附着反应的整体效率，以及对多功能病毒的严格EM测试 粒子平台成功的方法将应用于重要的蛋白质系统， 迄今为止还不能通过电子显微镜进行结构表征。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Knowledge of the three-dimensional structures of proteins and multi-protein assemblies is of great importance to the understanding of fundamental biological processes and the design of more effective Pharmaceuticals. For many such species, which cannot be crystallized and are too large for NMR analysis, electron microscopy (EM) is the only technique that can provide useful structural information. The attachment of electron-dense molecules, containing clusters of heavy metallic elements that scatter electrons with high efficiency, is often necessary to EM data collection and analysis. Practitioners of high-resolution EM usually have access to only two electron-dense materials and a limited set of attachment methods available from commercial suppliers. In many cases, these resources fail and the target must be either abandoned or laboriously re-engineered. We propose to create new electron-dense compounds and develop methods to rapidly identify the best ways to connect them to target proteins. The goal is to give the EM community an expanded set of materials and techniques for protein labeling, so that a greater fraction of targets can be structurally characterized. The effort will be comprised of equal parts chemical synthesis, chemical analysis to measure the overall efficiency of attachment reactions, and rigorous EM testing on a versatile virus particle platform. Successful methods will then be applied to important protein systems that have thus far resisted structural characterization by electron microscopy.