High‐resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages

Guo, Meng; Härtlová, Anetta; Dill, Brian D.; Prescott, Alan R.; Trost, Matthias

doi:10.1002/pmic.201400431

Cited by 74 publications

(57 citation statements)

References 31 publications

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“…Mass spectrometric analyses were conducted similarly as previously described (Dill et al , ; Guo et al , ). In detail, biological triplicates or quadruplicates of mixes of 1 μg of light‐labelled and 1 μg of heavy‐labelled samples were analysed on an Orbitrap Velos Pro mass spectrometer coupled to an Ultimate 3000 UHPLC system with a 50 cm Acclaim PepMap 100 or EasySpray analytical column (75 μm ID, 3 μm C18) in conjunction with a PepMap trapping column (100 μm × 2 cm, 5 μm C18; Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

Triggering MSR1 promotes JNK‐mediated inflammation in IL‐4‐activated macrophages

et al. 2019

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Alternatively activated M2 macrophages play an important role in maintenance of tissue homeostasis by scavenging dead cells, cell debris and lipoprotein aggregates via phagocytosis. Using proteomics, we investigated how alternative activation, driven by IL‐4, modulated the phagosomal proteome to control macrophage function. Our data indicate that alternative activation enhances homeostatic functions such as proteolysis, lipolysis and nutrient transport. Intriguingly, we identified the enhanced recruitment of the TAK1/MKK7/JNK signalling complex to phagosomes of IL‐4‐activated macrophages. The recruitment of this signalling complex was mediated through K63 polyubiquitylation of the macrophage scavenger receptor 1 (MSR1). Triggering of MSR1 in IL‐4‐activated macrophages leads to enhanced JNK activation, thereby promoting a phenotypic switch from an anti‐inflammatory to a pro‐inflammatory state, which was abolished upon MSR1 deletion or JNK inhibition. Moreover, MSR1 K63 polyubiquitylation correlated with the activation of JNK signalling in ovarian cancer tissue from human patients, suggesting that it may be relevant for macrophage phenotypic shift in vivo . Altogether, we identified that MSR1 signals through JNK via K63 polyubiquitylation and provides evidence for the receptor's involvement in macrophage polarization.

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Section: Methodsmentioning

confidence: 99%

Triggering MSR1 promotes JNK‐mediated inflammation in IL‐4‐activated macrophages

et al. 2019

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“…In addition, FcγR signaling by IgG [109], recognition of phosphatidylserine on apoptotic cells by TIM4 [110], and Dectin1 signaling by beads coupled to β-glucans and fungi 111, 112 also trigger the recruitment of LC3 to the phagosome. In addition, LC3 is also present on carboxylated latex-bead phagosomes [113], suggesting that LC3 recruitment is common to many phagosomes.LAP is at the interface between phagocytosis and autophagy, using both autophagy proteins, such as the class III PI3-kinase Vps34, and endosomal, LAP-specific proteins, such as Rubicon. Rubicon promotes PI3P production by Vps34 and ROS production by NOX2, which are both essential for LAP 109, 112.…”

Section: Impact Of Immune Signals On Phagosome Maturationmentioning

confidence: 99%

Patterns, Receptors, and Signals: Regulation of Phagosome Maturation

Pauwels

Trost

Beyaert

et al. 2017

Trends in Immunology

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Recognition of microbial pathogens and dead cells and their phagocytic uptake by specialized immune cells are essential to maintain host homeostasis. Phagosomes undergo fusion and fission events with endosomal and lysosomal compartments, a process called ‘phagosome maturation’, which leads to the degradation of the phagosomal content. However, many phagocytic cells also act as antigen-presenting cells and must balance degradation and peptide preservation. Emerging evidence indicates that receptor engagement by phagosomal cargo, as well as inflammatory mediators and cellular activation affect many aspects of phagosome maturation. Unsurprisingly, pathogens have developed strategies to hijack this machinery, thereby interfering with host immunity. Here, we highlight progress in this field, summarize findings on the impact of immune signals, and discuss consequences for pathogen elimination.

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“…For several pathogenic microorganisms, proteomic studies unraveled the specific protein composition of phagolysosomes for a better understanding of the phagosomal maturation process and its modulation by pathogens (15)(16)(17)(18)(19)(20)(21)(22)(23). These studies demonstrated that the protein composition of the phagosome is highly dynamic, specific for the ingested particle, depends on the stage of phagosomal maturation, the type or cell line and the activation status of the phagocyte.…”

Section: Introductionmentioning

confidence: 99%

Proteomics of Aspergillus fumigatus Conidia-containing Phagolysosomes Identifies Processes Governing Immune Evasion

Schmidt

Vlaic

Krüger

et al. 2018

Molecular & Cellular Proteomics

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Invasive infections by the human pathogenic fungus Aspergillus fumigatus start with the outgrowth of asexual, airborne spores (conidia) into the lung tissue of immunocompromised patients. The resident alveolar macrophages phagocytose conidia, which end up in phagolysosomes. However, A. fumigatus conidia resist phagocytic degradation to a certain degree. This is mainly attributable to the pigment 1,8-dihydroxynaphthalene (DHN) melanin located in the cell wall of conidia, which manipulates the phagolysosomal maturation and prevents their intracellular killing. To get insight in the underlying molecular mechanisms, we comparatively analyzed proteins of mouse macrophage phagolysosomes containing melanized wild-type (wt) or nonmelanized pksP mutant conidia. For this purpose, a protocol to isolate conidia-containing phagolysosomes was established and a reference protein map of phagolysosomes was generated. We identified 637 host and 22 A. fumigatus proteins that were differentially abundant in the phagolysosome. 472 of the host proteins were overrepresented in the pksP mutant and 165 in the wt conidia-containing phagolysosome. Eight of the fungal proteins were produced only in pksP mutant and 14 proteins in wt conidia-containing phagolysosomes. Bioinformatical analysis compiled a regulatory module, which indicates host processes affected by the fungus. These processes include vATPase-driven phagolysosomal acidification, Rab5 and Vamp8-dependent endocytic trafficking, signaling pathways, as well as recruitment of the Lamp1 phagolysosomal maturation marker and the lysosomal cysteine protease cathepsin Z. Western blotting and immunofluorescence analyses confirmed the proteome data and moreover showed differential abundance of the major metabolic regulator mTOR. Taken together, with the help of a protocol optimized to isolate A. fumigatus conidia-containing phagolysosomes and a potent bioinformatics algorithm, we were able to confirm A. fumigatus conidia-dependent modification of phagolysosomal processes that have been described before and beyond that, identify pathways that have not been implicated in A. fumigatus evasion strategy, yet.Mass spectrometry proteomics data are available via ProteomeXchange with identifiers PXD005724 and PXD006134.

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High‐resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow derived macrophages

Cited by 74 publications

References 31 publications

Triggering MSR1 promotes JNK‐mediated inflammation in IL‐4‐activated macrophages

Triggering MSR1 promotes JNK‐mediated inflammation in IL‐4‐activated macrophages

Patterns, Receptors, and Signals: Regulation of Phagosome Maturation

Proteomics of Aspergillus fumigatus Conidia-containing Phagolysosomes Identifies Processes Governing Immune Evasion

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