UBC Theses and Dissertations
Proteomic analysis of a dynamic Salmonella-host interactome Brown, Lyda Mimi
Salmonella is capable of evading the host immune response by secreting virulence factors (effectors) that enter the host and interfere with critical cell signaling networks. Although many of the individual effector proteins have been well characterized by traditional biochemical methods, a shift towards global strategies would offer a system’s view of the intricate network of interactions (interactome) between the host and pathogen. Protein profiling across size exclusion chromatography with stable isotope labeled amino acids in cell culture (PCP-SILAC) is a recent proteomic method developed to characterize the composition of protein complexes with the advantages of being quantitative, capable of monitoring dynamic changes, and being completely tag and chemical cross-link free. This method was applied to study the dynamic changes of host protein complexes as a result of Salmonella infection. Analysis of dynamic PCP-SILAC proteins led to the hypothesis that host translational machinery was being targeted by Salmonella during infection. To test this hypothesis, a fluorescent assay was used to measure protein synthesis of cells infected with WT Salmonella versus control (non-infected cells). Results from the protein synthesis analysis showed a decrease in host translation, supporting our hypothesis that Salmonella targets host translational machinery. I addition to this novel finding, this work provides a rich resource of candidate host proteins that may be involved with Salmonella’s pathogenesis.
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Attribution-NonCommercial-NoDerivs 2.5 Canada