Consortia for High-Throughput-Enabled Structural Biology Partnerships (U01)

描述(由申请人提供)：虽然结构生物学在研究致病分泌效应器方面很强大，但在很大程度上还没有被用于研究效应器与宿主蛋白的相互作用。在这项提议中，我们使用了已知和新的分泌效应器和一条实验管道，这将为模式病原体沙门氏菌如何颠覆宿主细胞功能提供广泛的见解。Aim 1同时使用广泛的分析方法提供对效应器蛋白质效力和功能的关键功能洞察，同时也是通过PSI网络将结构特征集中在最有价值的靶点上的重要选择步骤。AIM 2利用交联法和蛋白质组学提供了有关宿主蛋白和蛋白通路的有价值的信息，每个效应器与之相互作用，并为PSI网络提供了用于结构表征的宿主蛋白靶标的优先列表。目的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于PSI网络的结构表征。最终，对个体分泌效应物的结构-功能研究对宿主-病原体研究界来说是无价的。然而，从这种具有多个效应器的大型并行表征工作中获得的对宿主-病原体生物学的洞察将促进在更完整的系统水平上的理解。

DESCRIPTION (provided by applicant): 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 function. Aim 1 simultaneously provides key functional insights into effector protein potency and function using broad set of assays, while at the same time 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 characterization by PSI network. Ultimately, structure-function 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 characterization effort with multiple effectors will advance understanding at a more complete systems level.