Dataset Information

Profiling of Human Circulating Plasma Microparticle Associated Proteins by DDA and DIA

ABSTRACT: Cells of the vascular system release spherical vesicles, called microparticles (MP), in the size range of 0.1-1μm induced by a variety of stress factors resulting in variable MP concentrations between health and disease. Furthermore, MP have intercellular communication/signaling properties and interfere with inflammation and coagulation pathways. Today’s most used analytical technology for MP characterization, flow cytometry, is lacking sensitivity and specificity, which might have led to the publication of contradicting results in the past. We propose the use of nano-liquid chromatography coupled to two-stage mass spectrometry as a non-biased tool for quantitative MP proteome analysis. Using aliquots of 250 μL platelet-free plasma (PFP) from one individual donor, we achieved excellent inter-assay CV’s of 2.7 ± 1.7% (mean ± 1 S.D.) on individual peptide intensities across 27 data-dependent nanoLC-MS/MS runs performed over a period of 3.5 months. With quantitative proteomics, we show that MP composition between twelve volunteers were remarkably stable. MP protein composition is clearly distinguishable from whole cell lysates and we propose that this trait should be used as a quality criterion of MP purity. Furthermore, MP were damaged by freezing PFP. The damage was articulated by a loss of cytoplasm proteins, encompassing a specific set of proteins involved in regulating dynamic structures of the cytoskeleton, and thrombin activation leading to MP clotting. On the other hand, plasma membrane protein composition (cell markers) was not affected. Finally, we show that multiplexed data-independent acquisition can be used for relative quantification of target proteins using Skyline software.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Blood Plasma

DISEASE(S): Disease Free

SUBMITTER: Manfred Heller

LAB HEAD: Manfred Heller

PROVIDER: PXD003935 | Pride | 2016-10-18

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	14Jul01_MP1_90min_i01.raw	Raw
	14Jul01_MP1_90min_i02.raw	Raw
	14Jul01_MP1_90min_i03.raw	Raw
	14Jul11_MP2_90min_i01.raw	Raw
	14Jul11_MP2_90min_i02.raw	Raw

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Publications

Robust Label-free, Quantitative Profiling of Circulating Plasma Microparticle (MP) Associated Proteins.

Braga-Lagache Sophie S Buchs Natasha N Iacovache Mircea-Ioan MI Zuber Benoît B Jackson Christopher Benjamin CB Heller Manfred M

Molecular & cellular proteomics : MCP 20161012 12

Cells of the vascular system release spherical vesicles, called microparticles, in the size range of 0.1-1 μm induced by a variety of stress factors resulting in variable concentrations between health and disease. Furthermore, microparticles have intercellular communication/signaling properties and interfere with inflammation and coagulation pathways. Today's most used analytical technology for microparticle characterization, flow cytometry, is lacking sensitivity and specificity, which might ha ...[more]

PMID: 27738094

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