DETERMINING THE ARCHITECTURES OF MACROMOLECULAR ASSEMBLIES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 要了解活细胞的运作，我们需要知道其大分子组装的结构。在这里，我们展示了如何蛋白质组学数据可以用来确定这样的结构。该过程包括收集足够的和多样化的高质量数据，将这些数据转换为空间约束，以及使用约束来生成与数据一致的结构集合的优化。对集成的分析产生了组件的详细架构图。我们开发了一个具有挑战性的模型系统，核孔复合体（NPC）的方法。NPC充当动态屏障，控制进出细胞核，并且在酵母中是456个蛋白质的50 MDa组装体。由此产生的结构，在随附的论文中提出，揭示了NPC中蛋白质的配置，提供了对其进化和结构原理的见解。本方法应该适用于许多其他大分子组装体。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To understand the workings of a living cell, we need to know the architectures of its macromolecular assemblies. Here we show how proteomic data can be used to determine such structures. The process involves the collection of sufficient and diverse high-quality data, translation of these data into spatial restraints, and an optimization that uses the restraints to generate an ensemble of structures consistent with the data. Analysis of the ensemble produces a detailed architectural map of the assembly. We developed our approach on a challenging model system, the nuclear pore complex (NPC). The NPC acts as a dynamic barrier, controlling access to and from the nucleus, and in yeast is a 50 MDa assembly of 456 proteins. The resulting structure, presented in an accompanying paper, reveals the configuration of the proteins in the NPC, providing insights into its evolution and architectural principles. The present approach should be applicable to many other macromolecular assemblies.