| Quantitative proteomics reveals that only a subset of the endoplasmic reticulum contributes to the phagosome. | |
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MedLine Citation:
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PMID: 22427703 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Phagosomes, by killing and degrading pathogens for antigen presentation, are organelles implicated in key aspects of innate and adaptive immunity. Although it has been well established that phagosomes consist of membranes from the plasma membrane, endosomes, and lysosomes, the notion that the endoplasmic reticulum (ER) membrane could play an important role in the formation of the phagosome is debated. However, a method to accurately estimate the contribution of potential source organelles and contaminants to the phagosome proteome has been lacking. Herein, we have developed a proteomic approach for objectively quantifying the contribution of various organelles to the early and late phagosomes by comparing these fractions to their total membrane and postnuclear supernatant of origin in the J774A.1 murine macrophage cell line. Using quantitative label-free mass spectrometry, the abundance of peptides corresponding to hundreds of proteins was estimated and attributed to one of five organelles (e.g. plasma membrane, endosomes/lysosomes, ER, Golgi, and mitochondria). These data in combination with a stable isotope labeling in cell culture method designed to detect potential contaminant sources revealed that the ER is part of the phagosomal membrane and contributes ≈ 20% of the early phagosome proteome. In addition, only a subset of ER proteins is recruited to the phagosome, suggesting that a specific subdomain(s) of the ER might be involved in phagocytosis. Western blotting and immunofluorescence substantially validated this conclusion; we were able to demonstrate that the fraction of the ER in which the ER marker GFP-KDEL accumulates is excluded from the phagosomes, whereas that containing the mVenus-Syntaxin 18 is recruited. These results highlight promising new avenues for the description of the pathogenic mechanisms used by Leishmania, Brucella, and Legionella spp., which thrive in ER-rich phagosomes. |
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Authors:
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François-Xavier Campbell-Valois; Matthias Trost; Magali Chemali; Brian D Dill; Annie Laplante; Sophie Duclos; Shayan Sadeghi; Christiane Rondeau; Isabel C Morrow; Christina Bell; Etienne Gagnon; Kiyokata Hatsuzawa; Pierre Thibault; Michel Desjardins |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2012-03-15 |
Journal Detail:
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Title: Molecular & cellular proteomics : MCP Volume: 11 ISSN: 1535-9484 ISO Abbreviation: Mol. Cell Proteomics Publication Date: 2012 Jul |
Date Detail:
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Created Date: 2012-07-09 Completed Date: 2012-10-30 Revised Date: 2013-04-16 |
Medline Journal Info:
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Nlm Unique ID: 101125647 Medline TA: Mol Cell Proteomics Country: United States |
Other Details:
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Languages: eng Pagination: M111.016378 Citation Subset: IM |
Affiliation:
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Département de Pathologie et Biologie Cellulaire, Université de Montréal, Montréal, Quebec, H3C 3J7, Canada. fxcamval@pasteur.fr |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Biological Markers / analysis Blotting, Western Cell Line Cell Membrane / chemistry, metabolism, ultrastructure Endoplasmic Reticulum / chemistry*, metabolism, ultrastructure Endosomes / chemistry, metabolism, ultrastructure Fluorescent Antibody Technique Golgi Apparatus / chemistry, metabolism, ultrastructure Isotope Labeling Lysosomes / chemistry, metabolism, ultrastructure Macrophages / cytology, metabolism*, ultrastructure Mass Spectrometry Mice Oligopeptides Phagocytosis Phagosomes / chemistry*, metabolism, ultrastructure Plasmids Protein Sorting Signals Proteomics / methods* Qa-SNARE Proteins Transfection |
| Grant Support | |
ID/Acronym/Agency:
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//Canadian Institutes of Health Research; //Medical Research Council |
| Chemical | |
Reg. No./Substance:
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0/Biological Markers; 0/Oligopeptides; 0/Protein Sorting Signals; 0/Qa-SNARE Proteins; 113516-56-6/lysyl-aspartyl-glutamyl-leucine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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