In vivo Proteomics Approaches for the Analysis of Bacterial Adaptation Reactions in Host–Pathogen Settings
Proteome profiling of bacteria internalized by host cells is still a challenging task, due to low amounts of bacterial proteins in host–pathogen settings and the high amounts of contaminating host proteins. Here, we describe a workflow for the enrichment of intracellular bacteria by fluorescence activated cell sorting which in combination with highly sensitive LC-MS/MS allows monitoring of about 1200 proteins from 2 to 4 × 106 internalized bacterial cells as starting material.
Key wordsHost–pathogen interaction Staphylococcus aureus Intracellular bacteria Epithelial cells Internalization Bacterial cell sorting Mass spectrometry Proteomics
This work was supported by the German Research Foundation Grant SFB/TR34.