Document Detail


Pathogen Vacuole Purification from Legionella-Infected Amoeba and Macrophages.
MedLine Citation:
PMID:  23150404     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Legionella pneumophila replicates intracellularly in environmental and immune phagocytes within a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). Formation of LCVs is strictly dependent on the Icm/Dot type IV secretion system and the translocation of "effector" proteins into the cell. Some effector proteins decorate the LCV membrane and subvert host cell vesicle trafficking pathways. Here we describe a method to purify intact LCVs from Dictyostelium discoideum amoebae and RAW 264.7 murine macrophages. The method comprises a two-step protocol: first, LCVs are enriched by immuno-magnetic separation using an antibody against a bacterial effector protein specifically localizing to the LCV membrane, and second, the LCVs are further purified by density gradient centrifugation. The purified LCVs can be characterized by proteomics and other biochemical approaches.
Authors:
Christine Hoffmann; Ivo Finsel; Hubert Hilbi
Related Documents :
23835174 - Screening for glycosylphosphatidylinositol (gpi)-modified cell wall proteins in pichia ...
23919454 - A novel fluorescent assay and catalytic properties of crh1 and crh2 yeast cell wall tra...
23931504 - Fluorescent protein applications in microscopy.
11535614 - A new view of the spindle checkpoint.
17689064 - Ubiquitin-dependent sorting of integral membrane proteins for degradation in lysosomes.
19028884 - Oryr is a luxr-family protein involved in interkingdom signaling between pathogenic xan...
Publication Detail:
Type:  JOURNAL ARTICLE    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  954     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  309-321     Citation Subset:  -    
Affiliation:
Max von Pettenkofer Institute, Ludwig-Maximilians University, Munich, Germany.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Electron Microscopy of Legionella and Legionella-Infected Cells.
Next Document:  Methods for determining protein translocation by the Legionella pneumophila Dot/Icm type IV secretio...