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Front Cell Infect Microbiol. 2015 Jun 2;5:48. doi: 10.3389/fcimb.2015.00048. eCollection 2015.

Purification and proteomics of pathogen-modified vacuoles and membranes.

Author information

1
Chair of Microbiology, Biocenter, University of Würzburg Würzburg, Germany.
2
Division of Microbiology, University of Osnabrück Osnabrück, Germany.
3
Institute of Microbiology, Ernst-Moritz-Arndt University Greifswald Greifswald, Germany.
4
Priority Area Infections, Cellular Microbiology, Research Center Borstel, Leibniz Center for Medicine and Biosciences Borstel, Germany.
5
Department of Medicine, Max von Pettenkofer Institute, Ludwig-Maximilians University Munich Munich, Germany ; Department of Medicine, Institute of Medical Microbiology, University of Zürich Zürich, Switzerland.

Abstract

Certain pathogenic bacteria adopt an intracellular lifestyle and proliferate in eukaryotic host cells. The intracellular niche protects the bacteria from cellular and humoral components of the mammalian immune system, and at the same time, allows the bacteria to gain access to otherwise restricted nutrient sources. Yet, intracellular protection and access to nutrients comes with a price, i.e., the bacteria need to overcome cell-autonomous defense mechanisms, such as the bactericidal endocytic pathway. While a few bacteria rupture the early phagosome and escape into the host cytoplasm, most intracellular pathogens form a distinct, degradation-resistant and replication-permissive membranous compartment. Intracellular bacteria that form unique pathogen vacuoles include Legionella, Mycobacterium, Chlamydia, Simkania, and Salmonella species. In order to understand the formation of these pathogen niches on a global scale and in a comprehensive and quantitative manner, an inventory of compartment-associated host factors is required. To this end, the intact pathogen compartments need to be isolated, purified and biochemically characterized. Here, we review recent progress on the isolation and purification of pathogen-modified vacuoles and membranes, as well as their proteomic characterization by mass spectrometry and different validation approaches. These studies provide the basis for further investigations on the specific mechanisms of pathogen-driven compartment formation.

KEYWORDS:

Chlamydia; Legionella; Mycobacterium; Salmonella; Simkania; host-pathogen interactions; immuno-magnetic purification; pathogen vacuole

PMID:
26082896
PMCID:
PMC4451638
DOI:
10.3389/fcimb.2015.00048
[Indexed for MEDLINE]
Free PMC Article

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