Consortia for High-Throughput-Enabled Structural Biology Partnerships

虽然结构生物学在研究致病分泌效应物方面很强大，但还没有在很大程度上应用于 研究效应器-宿主蛋白相互作用的程度。在这个提案中，我们使用了已知的和新的秘密 效应器和实验管道将提供对模型病原体如何。 沙门氏菌，颠覆宿主细胞功能。AIM 1同时提供对效应器的关键功能洞察 蛋白质的效力和功能使用广泛的分析，同时FIME是一个重要的 选择步骤，通过PSI网络将结构表征集中在最有价值的目标上。目标2 提供有关宿主蛋白质和蛋白质途径的有价值的信息，使用交联剂和 蛋白质组学，每个效应器与之相互作用，并向PSI网络提供按优先顺序排列的宿主列表 用于结构表征的蛋白质靶标。目标3询问可能的特定蛋白质-蛋白质 相互作用，并提供一系列有效的蛋白质-蛋白质相互作用和可能的配体 用PSI网络进行结构特征分析。最终，对个体分泌物的结构-功能研究 效应器对于宿主-病原体研究界来说是无价的。然而，对寄主病原体的洞察 从这种具有多个效应器的大规模并行特性努力中获得的生物学将会取得进展 在更完整的系统层面上理解。

While powerful for studying pathogenic secreted effectors, structural biology has not been used to a large extent to study effector-host protein interactions. In this proposal, we use known and novel secreted effectors and an experimental pipeline that will provide broad insights into how a model pathogen. Salmonella, subverts host cell funcfion. Aim 1 simultaneously provides key functional insights into effector protein potency and funcfion using broad set of assays, while at the same fime serves as an important selection step to focus structural characterization by the PSI network on the most valuable targets. Aim 2 provides valuable information about the host proteins and protein pathways, using cross-linking and proteomics, that each effector interacts with and provides the PSI network with a prioritized list of host protein targets for structural characterization. Aim 3 interrogates possible specific protein-protein interactions and provides a selection of validated protein-protein interactions and possibly ligands for structural characterizafion by PSI network. Ultimately, structure-funcfion studies of individual secreted effectors are invaluable to the host-pathogen research community. However, the insights into host-pathogen biology gained from this large parallel characterizafion effort with multiple effectors will advance understanding at a more complete systems level.