Dataset Information

Platelet Activation Results in Modulation of miRNA, mRNA, and Protein Expression Profiles: New Insights into the Pathophysiology of Platelet Activation

ABSTRACT: Leucocytes-decontaminated platelet-rich plasma (PRP) from 15 healthy volunteers was collected and activated with collagen, or Thrombin Activator Receptor Peptide (TRAP). Total protein extracts (75 microg protein/sample), coming from two biological replicates, were precipitated using 2-D Clean Up Kit (Amersham Bioscience). Total protein extracts were precipitated and dissolved into 20 microl of iTRAQ dissolution buffer. Protein alkylation, trypsin digestion and labeling of the resulting peptides were performed according to manufacturer’s instructions (Applied Biosystems) followed by strong cation exchange fractionation and LC-MS/MS analysis of the peptides as previously described (Lietzén et al, PLoS Pathogens). Protein identification and relative quantitation were performed with Paragon search algorithm using ProteinPilot 4 interface (Applied Biosystems). Database searches were performed against human protein sequences in UniProt database (version 120813 with 20231 human entries). The search criteria were: cysteine alkylation with MMTS, trypsin digestion, biological modifications allowed, thorough search and detected protein threshold of 95% confidence (Paragon Unused ProtScore > 1.3). Additionally, automatic bias correction was used to correct for uneven protein loading.

INSTRUMENT(S): QSTAR

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Tuula Nyman

PROVIDER: PXD000621 | Pride | 2015-04-27

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	130619GiovanniiTRAQ1B12.wiff	Wiff
	130619GiovanniiTRAQ1B12.wiff.scan	Wiff
	130619GiovanniiTRAQ1B5.wiff	Wiff
	130619GiovanniiTRAQ1B5.wiff.scan	Wiff
	130619GiovanniiTRAQ1B9.wiff	Wiff

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Publications

Activating stimuli induce platelet microRNA modulation and proteome reorganisation.

Cimmino Giovanni G Tarallo Roberta R Nassa Giovanni G De Filippo Maria Rosaria MR Giurato Giorgio G Ravo Maria M Rizzo Francesca F Conte Stefano S Pellegrino Grazia G Cirillo Plinio P Calabro Paolo P Öhman Tiina T Nyman Tuula A TA Weisz Alessandro A Golino Paolo P

Thrombosis and haemostasis 20150423 1

Platelets carry megakaryocyte-derived mRNAs whose translation efficiency before and during activation is not known, although this can greatly affect platelet functions, both under basal conditions and in response to physiological and pathological stimuli, such as those involved in acute coronary syndromes. Aim of the present study was to determine whether changes in microRNA (miRNA) expression occur in response to activating stimuli and whether this affects activity and composition of platelet t ...[more]

PMID: 25903651

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