MassIVE MSV000080769

Imported Reanalysis Dataset Public PXD005083

Proteomic and bioinformatic characterization of proteins secreted conventionally and unconventionally from human primary macrophages upon LPS transfection

Description

The non-canonical caspase-4/5 inflammasome has been only very recently characterized. It is expressed by macrophages, the principal effector cells of the innate immunity system, and is activated by gram-negative bacteria, ER stress, and UV radiation. Activation of this inflammasome results in pyroptosis, inflammatory cell death, and secretion of pro-inflammatory cytokines of the interleukin-1 family. In our model we transfected human primary macrophages with Lipopolysaccharide, a cell wall component of gram-negative bacteria, to simulate bacteria entering the cell and thus activating the caspase-4/5 inflammasome. We then performed high-throughput proteomics to identify proteins secreted during the stimulation. The secretome was separated into two fractions using size-exclusion centrifugation, with a 100 kDa cut-off we enriched extracellular vesicles (EVs) and the flow through was concentrated over 10 kDa to yield the rest-secretome (RS) samples. The EV and RS samples of LPS transfected cells were then compared to untreated cells. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Macrophages ; non-canonical inflammasome ; LPS ; Innate immunity

Contact

Principal Investigators:
(in alphabetical order)
Tuula Nyman, Head of Proteomics Core Facility, Rikshospitalet Oslo, Department of Immunology, University of Norway, Norway, N/A
Submitting User: ccms

Publications

Lorey MB, Rossi K, Eklund KK, Nyman TA, Matikainen S.
Global characterization of protein secretion from human macrophages following non-canonical caspase-4/5 inflammasome activation.
Mol. Cell Proteomics. Epub 2017 Mar 14.

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Experimental Design
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Identification Results
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Quantification Results
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Number of distinct conditions across all analyses (original submission and reanalyses) associated with this dataset.

Distinct condition labels are counted across all files submitted in the "Metadata" category having a "Condition" column in this dataset.

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Number of distinct biological replicates across all analyses (original submission and reanalyses) associated with this dataset.

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Number of distinct technical replicates across all analyses (original submission and reanalyses) associated with this dataset.

The technical replicate count is defined as the maximum number of times any one distinct combination of condition and biological replicate was analyzed across all files submitted in the "Metadata" category. In the case of fractionated experiments, only the first fraction is considered.

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Originally identified proteins that were automatically remapped by MassIVE to proteins in the SwissProt human reference database.

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Number of distinct protein accessions reported across all analyses (original submission and reanalyses) associated with this dataset.

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Number of distinct unmodified peptide sequences reported across all analyses (original submission and reanalyses) associated with this dataset.

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Number of distinct peptide sequences (including modified variants or peptidoforms) reported across all analyses (original submission and reanalyses) associated with this dataset.

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Total number of peptide-spectrum matches (i.e. spectrum identifications) reported across all analyses (original submission and reanalyses) associated with this dataset.

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Number of distinct proteins quantified across all analyses (original submission and reanalyses) associated with this dataset.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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Number of distinct proteins found to be differentially abundant in at least one comparison across all analyses (original submission and reanalyses) associated with this dataset.

A protein is differentially abundant if its change in abundance across conditions is found to be statistically significant with an adjusted p-value <= 0.05 and lists no issues associated with statistical tests for differential abundance.

Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

"N/A" means no results of this type were submitted.
This dataset may not contain all raw spectra data as originally deposited in PRIDE. It has been imported to MassIVE for reanalysis purposes, so its spectra data here may consist solely of processed peak lists suitable for reanalysis with most software.