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Purification and proteomics of pathogen-modified vacuoles and membranes

Please always quote using this URN: urn:nbn:de:bvb:20-opus-151823
  • 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 earlyCertain 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.show moreshow less

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Metadaten
Author: Jo-Ana Herweg, Nicole Hansmeier, Andreas Otto, Anna C. Geffken, Prema Subbarayal, Bhupesh K. Prusty, Dörte Becher, Michael Hensel, Ulrich E. Schaible, Thomas Rudel, Hubert Hilbi
URN:urn:nbn:de:bvb:20-opus-151823
Document Type:Journal article
Faculties:Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Frontiers in Cellular and Infection Microbiology
Year of Completion:2015
Volume:5
Issue:48
Source:Frontiers in Cellular and Infection Microbiology 5, 48 (2015). DOI: 10.3389/fcimb.2015.00048
DOI:https://doi.org/10.3389/fcimb.2015.00048
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Tag:Chlamydia; Chlamydia trachomatis; III secretion system; Legionella; Mycobacterium; Mycobacterium tuberculosis; Salmonella; Simkania; Simkania negevensis; bacterium Legionella pneumophila; endocytic multivesicular bodies; endoplasmic reticulum; host pathogen interactions; immuno-magnetic purification; intracellular bacteria; pathogen vacuole; phagosome maturation arrest; spectrometry-based proteomics
Release Date:2017/10/27
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International