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.
Advanced mass spectrometry methods with structure determinations will allow for improved understanding of the interactions between human host proteins and pathogens;this research may lead to new therapies for infectious diseases. The specific pathogen model to be studied here is Salmonella, a pathogen that has been the cause of a number of significant food-borne outbreaks in recent years.
|Eletsky, Alexander; Michalska, Karolina; Houliston, Scott et al. (2014) Structural and functional characterization of DUF1471 domains of Salmonella proteins SrfN, YdgH/SssB, and YahO. PLoS One 9:e101787|
|Lu, Jun; Wu, Ruiying; Adkins, Joshua N et al. (2014) Crystal structures of the F and pSLT plasmid TraJ N-terminal regions reveal similar homodimeric PAS folds with functional interchangeability. Biochemistry 53:5810-9|
|Merkley, Eric D; Rysavy, Steven; Kahraman, Abdullah et al. (2014) Distance restraints from crosslinking mass spectrometry: mining a molecular dynamics simulation database to evaluate lysine-lysine distances. Protein Sci 23:747-59|
|Nakayasu, Ernesto S; Tempel, Rebecca; Cambronne, Xiaolu A et al. (2013) Comparative phosphoproteomics reveals components of host cell invasion and post-transcriptional regulation during Francisella infection. Mol Cell Proteomics 12:3297-309|
|Niemann, George S; Brown, Roslyn N; Mushamiri, Ivy T et al. (2013) RNA type III secretion signals that require Hfq. J Bacteriol 195:2119-25|
|Nakayasu, Ernesto S; Ansong, Charles; Brown, Joseph N et al. (2013) Evaluation of selected binding domains for the analysis of ubiquitinated proteomes. J Am Soc Mass Spectrom 24:1214-23|
|Merkley, Eric D; Cort, John R; Adkins, Joshua N (2013) Cross-linking and mass spectrometry methodologies to facilitate structural biology: finding a path through the maze. J Struct Funct Genomics 14:77-90|
|Nakayasu, Ernesto S; Brown, Roslyn N; Ansong, Charles et al. (2013) Multi-omic data integration links deleted in breast cancer 1 (DBC1) degradation to chromatin remodeling in inflammatory response. Mol Cell Proteomics 12:2136-47|
|Ansong, Charles; Wu, Si; Meng, Da et al. (2013) Top-down proteomics reveals a unique protein S-thiolation switch in Salmonella Typhimurium in response to infection-like conditions. Proc Natl Acad Sci U S A 110:10153-8|
|Michalska, Karolina; Brown, Roslyn N; Li, Hui et al. (2013) New sub-family of lysozyme-like proteins shows no catalytic activity: crystallographic and biochemical study of STM3605 protein from Salmonella Typhimurium. J Struct Funct Genomics 14:1-10|
Showing the most recent 10 out of 12 publications