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

描述（由申请人提供）：虽然结构生物学对于研究致病性分泌效应物很有效，但尚未在很大程度上用于研究效应物-宿主蛋白质相互作用。在这项提案中，我们使用已知的和新的分泌效应物和实验管道，这将提供广泛的见解，模型病原体，沙门氏菌，颠覆宿主细胞功能。目标1同时提供了使用广泛的测定集对效应蛋白效力和功能的关键功能见解，同时作为重要的选择步骤，通过PSI网络将结构表征集中在最有价值的靶标上。 目的2提供了有价值的信息，宿主蛋白质和蛋白质通路，使用交联和蛋白质组学，每个效应器相互作用，并提供了PSI网络与宿主蛋白质靶点的优先级列表的结构表征。目的3探讨可能的特异性蛋白质-蛋白质相互作用，并为PSI网络的结构表征提供一系列经过验证的蛋白质-蛋白质相互作用和可能的配体。最终，个体分泌效应物的结构-功能研究对宿主-病原体研究界是无价的。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-病原体生物学的见解将在更完整的系统水平上推进理解。

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.