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In vivo Proteomics Approaches for the Analysis of Bacterial Adaptation Reactions in Host–Pathogen Settings

  • Henrike Pförtner
  • Maren Depke
  • Kristin Surmann
  • Frank Schmidt
  • Uwe Völker
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1841)

Abstract

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 words

Host–pathogen interaction Staphylococcus aureus Intracellular bacteria Epithelial cells Internalization Bacterial cell sorting Mass spectrometry Proteomics 

Notes

Acknowledgments

This work was supported by the German Research Foundation Grant SFB/TR34.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Henrike Pförtner
    • 1
  • Maren Depke
    • 1
  • Kristin Surmann
    • 1
  • Frank Schmidt
    • 2
  • Uwe Völker
    • 2
  1. 1.Interfaculty Institute for Genetics and Functional GenomicsUniversity Medicine GreifswaldGreifswaldGermany
  2. 2.Interfaculty Institute for Genetics and Functional GenomicsUniversity Medicine GreifswaldGreifswaldGermany

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