Dataset Information

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

ABSTRACT: 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.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Macrophage

SUBMITTER: Tuula Nyman

LAB HEAD: Tuula Nyman

PROVIDER: PXD005083 | Pride | 2017-03-07

REPOSITORIES: Pride

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Publications

Global Characterization of Protein Secretion from Human Macrophages Following Non-canonical Caspase-4/5 Inflammasome Activation.

Lorey Martina B MB Rossi Katriina K Eklund Kari K KK Nyman Tuula A TA Matikainen Sampsa S

Molecular & cellular proteomics : MCP 20170214 4 suppl 1

Gram-negative bacteria are associated with a wide spectrum of infectious diseases in humans. Inflammasomes are cytosolic protein complexes that are assembled when the cell encounters pathogens or other harmful agents. The non-canonical caspase-4/5 inflammasome is activated by Gram-negative bacteria-derived lipopolysaccharide (LPS) and by endogenous oxidized phospholipids. Protein secretion is a critical component of the innate immune response. Here, we have used label-free quantitative proteomic ...[more]

PMID: 28196878

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