Dataset Information

Characterization of the proteome of human blood platelet sercretory granules using gas-phase fractionation (GPF)

ABSTRACT: We performed an extensive qualitative characterization of the platelet granule proteome using subcellular fractionation followed by mass spectrometry analysis and functional annotation. Platelets were isolated from whole blood by centrifugation and their level of purity was tested by microscopic inspection, western blot and Leucocount kit. Subcellular fractionation based on sucrose gradient was used to enrich sample in secretory granules. Then sample quality was assessed by western blot and electronic microscopy, prior proteomic study. Samples were trypsin digested and analyzed using an Orbitrap Velos in a gas-phase fractionation mode. The m/z ranges for the selection of precursor ions were 400-521, 516-690, 685-986 and 963-2000 Thomsons. The 2 technical replicates were analyzed by a GPF series of injections. Protein identification was performed using the EasyProt resource. The module EasyProtConv was used to generate the MGF file from the raw data. The peaklist files were searched against the UniProt database (2011_02 of 08-Feb-2011) using EasyProt. Eight-hundred-and-twenty-seven proteins were identified, most of them being associated to granules and to the granule’s secretory machinery.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

SUBMITTER: Pierre Fontana

LAB HEAD: Pierre Fontana

PROVIDER: PXD000618 | Pride | 2014-03-06

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	2011_12_14_ALS_OT_101_ANZ_LB2_7_1.acc.mgf	Mgf
	2011_12_14_ALS_OT_101_ANZ_LB2_7_1.raw	Raw
	2011_12_14_ALS_OT_102_ANZ_LB2_7_2.acc.mgf	Mgf
	2011_12_14_ALS_OT_102_ANZ_LB2_7_2.raw	Raw
	2011_12_14_ALS_OT_103_ANZ_LB2_7_3.acc.mgf	Mgf

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Publications

Characterization of the platelet granule proteome: evidence of the presence of MHC1 in alpha-granules.

Zufferey Anne A Schvartz Domitille D Nolli Séverine S Reny Jean-Luc JL Sanchez Jean-Charles JC Fontana Pierre P

Journal of proteomics 20140216

In the present study, we performed an extensive qualitative characterization of the platelet granule proteome using subcellular fractionation followed by mass spectrometry analysis and functional annotation. Eight-hundred-and-twenty-seven proteins were identified, most of them being associated to granules and to the granule's secretory machinery. Functional pathway analysis revealed 30 pathways, including the major histocompatibility complex class 1 (MHC I) presenting antigen pathway. This pathw ...[more]

PMID: 24549006

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