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Legionella pp 445-464 | Cite as

Subcellular Protein Fractionation in Legionella pneumophila and Preparation of the Derived Sub-proteomes for Analysis by Mass Spectrometry

  • Sandra Maaß
  • Gina Moog
  • Dörte BecherEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)

Abstract

Classical proteomic techniques are perfectly suited to reflect changes in the metabolism by detection of changed protein synthesis rates and protein abundances in a global protein-centered analysis. Although the proteome of microbes is considered as rather low complex, usually the subcellular fractionation of proteins leads to higher proteome coverage which might be important for the proteome quantification. Additionally, such fractionation provides the possibility to detect changes in the protein localization as well as the protein abundance in single sub-proteomes. Here, a workflow for subcellular fractionation of Legionella pneumophila into cytosolic, periplasmic, membrane, and extracellular proteins for global proteome analyses is provided. The methods included in this workflow can be used to analyze the adaptation of L. pneumophila to different environmental and nutritional situations during infection or during different life cycle stages including planktonic or biofilm phase.

Key words

Proteomics Protein Protein enrichment Protein identification Subcellular localization Virulence factors Metabolic enzymes 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Microbiology, Center for Functional Genomics of MicrobesUniversity of GreifswaldGreifswaldGermany

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