Dataset Information

Protein ADP-ribosylation, phosphorylation, and interactome dynamics in macrophage activation.

ABSTRACT: The innate immune response relies on efficient, robust and fast protein signaling networks to relay information related to pathogen or viral detection. This communication is mediated primarily through protein-protein interactions and post-translational modifications (PTMs), events which are best characterized by mass spectrometry (MS)-based proteomics. This in-depth study uses MS to identify changes in protein signaling networks of Lipopolysaccharide (LPS)-stimulated human and mouse macrophages, at the level of single PTMs (via phosphorylation and ADP-ribosylation site ID) and protein complexes (via size exclusion chromatography and immunoprecipitation). The result is a curated meta-database of 6,475 proteins including 2,311 ADP-ribosylated proteins and 2,284 phosphoproteins present in LPS-stimulated macrophages. Follow up studies characterized the ASK protein complex – which appeared to dissociate upon LPS stimulation – and a complex which formed upon LPS stimulation and contained the poly(ADP-ribosyl) transferase PARP9 protein.

INSTRUMENT(S): Q Exactive HF, Velos Plus

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Bone Marrow, Blood, Macrophage

SUBMITTER: Casey Daniels

LAB HEAD: Aleksandra Nita-Lazar

PROVIDER: PXD016469 | Pride | 2020-06-17

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	160316_CMD_Raw_E2_15min.raw	Raw
	160316_CMD_Raw_E2_30min.raw	Raw
	160316_CMD_Raw_E2_5min.raw	Raw
	160316_CMD_Raw_E2_60min.raw	Raw
	160316_CMD_Raw_E2_untreated.raw	Raw

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Publications

Dynamic ADP-Ribosylome, Phosphoproteome, and Interactome in LPS-Activated Macrophages.

Daniels Casey M CM Kaplan Pauline R PR Bishof Isaac I Bradfield Clinton C Tucholski Trisha T Nuccio Arthur G AG Manes Nathan P NP Katz Samuel S Fraser Iain D C IDC Nita-Lazar Aleksandra A

Journal of proteome research 20200701 9

We have used mass spectrometry (MS) to characterize protein signaling in lipopolysaccharide (LPS)-stimulated macrophages from human blood, human THP1 cells, mouse bone marrow, and mouse Raw264.7 cells. Protein ADP-ribosylation was truncated down to phosphoribose, allowing for enrichment and identification of the resulting phosphoribosylated peptides alongside phosphopeptides. Size exclusion chromatography-MS (SEC-MS) was used to separate proteoforms by size; protein complexes were then identifie ...[more]

PMID: 32529831

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