title: High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages.
abstract: Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in-depth proteomics characterization of phagosomes from RAW 264.7 and bone marrow derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicate that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec-1. Moreover, bone marrow derived macrophages phagosomes mature considerably faster by fusion with endosomes and the lysosome which we validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. All MS data have been deposited in the ProteomeXchange with identifier PXD001293 (http://proteomecentral.proteomexchange.org/dataset/PXD001293).
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Science, University of Dundee, Scotland, UK.
authors.forename: Manman
authors.initials: M
authors.lastname: Guo
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Science, University of Dundee, Scotland, UK.
authors.forename: Anetta
authors.initials: A
authors.lastname: Härtlova
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Science, University of Dundee, Scotland, UK.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Dill
authors.affiliation: Division of Cell Signalling and Immunology, College of Life Science, University of Dundee, Scotland, UK.
authors.forename: Alan R
authors.initials: AR
authors.lastname: Prescott
authors.affiliation: Division of Computational Biology, College of Life Sciences, University of Dundee, Scotland, UK.
authors.forename: Marek
authors.initials: M
authors.lastname: Gierliński
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Science, University of Dundee, Scotland, UK.
authors.forename: Matthias
authors.identifier: http://orcid.org/0000-0002-5732-700X
authors.initials: M
authors.lastname: Trost
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D030562:Databases, Protein; D008264:Macrophages / Q000737:chemistry*; D051379:Mice; D010588:Phagosomes / Q000737:chemistry*; D060066:Protein Interaction Maps; D020543:Proteome / Q000032:analysis / Q000737:chemistry / Q000145:classification; D040901:Proteomics; D000067996:RAW 264.7 Cells / Q000737:chemistry / Q000166:cytology
chemical_list: D020543:Proteome
keywords: Cell biology; Macrophage; Phagosome; Proteomics RAW 264.7