Abstract
Platelets are a heterogeneous small anucleate blood cell population with a central role both in physiological haemostasis and in pathological states, spanning from thrombosis to inflammation, and cancer. Recent advances in proteomic studies provided additional important information concerning the platelet biology and the response of platelets to several pathophysiological pathways. Platelets circulate systemically and can be easily isolated from human samples, making proteomic application very interesting for characterizing the complexity of platelet functions in health and disease as well as for identifying and quantifying potential platelet proteins as biomarkers and novel antiplatelet therapeutic targets. To date, the highly dynamic protein content of platelets has been studied in resting and activated platelets, and several subproteomes have been characterized including platelet-derived microparticles, platelet granules, platelet releasates, platelet membrane proteins, and specific platelet post-translational modifications. In this review, a critical overview is provided on principal platelet proteomic studies focused on platelet biology from signaling to granules content, platelet proteome changes in several diseases, and the impact of drugs on platelet functions. Moreover, recent advances in quantitative platelet proteomics are discussed, emphasizing the importance of targeted quantification methods for more precise, robust and accurate quantification of selected proteins, which might be used as biomarkers for disease diagnosis, prognosis and therapy, and their strong clinical impact in the near future.
Highlights
Over the past twenty years, mass spectrometry (MS) and its application in proteomic studies lead to the compilation of an extensive list of proteins expressed in platelets and relevant data on protein–protein interactions and post-translational modifications (PTMs)
* differentially expressed proteins are reported between brackets; 1-DE, 1-dimensional electrophoresis; 2D-DIGE, 2- dimensional differential gel electrophoresis; 2-DE, 2-dimensional electrophoresis; 4-HNE, 4-hydroxynonenale; acute coronary syndromes (ACS), acute coronary syndrome; AD, Alzheimer’s disease; ASA, acetyl salicylic acid; ChaFRADIC, charge-based fractional diagonal chromatography; COFRADIC, combined fractional diagonal chromatography; IB, immunoblotting; IHC, immunohistochemistry; IMAC, Immobilized metal affinity chromatography; IP, immunoprecipitation; LC, liquid chromatography; MALDI, matrix-assisted laser desorption ionization; MRM, multiple reaction monitoring; MS, mass spectrometry; MudPIT, Multidimensional Protein Identification Technology; NSTEMI, non ST-elevation myocardial infarction; PCI, Percutaneous coronary intervention; PRM, parallel reaction monitoring; SCAD, stable coronary artery disease; SCX, strong cation exchange; STEMI, ST-elevation myocardial infarction; TOF, time of flight
García et al performed a differential proteome analysis of intracellular signaling cascades using 2-DE coupled with nanoLC-MS intensities or tandem mass spectrometry (MS/MS) in human platelets stimulated with thrombin-receptor activated peptide (TRAP), which is a synthetic peptide with full agonist properties for thrombin receptor protease-activated receptors (PAR)-1 activation [98]
Summary
Platelets are small anucleate blood cells produced by megakaryocytes in the bone marrow and lungs [1,2]. Platelets are highly specialized effector cells in physiological haemostasis and play a central role in pathological thrombosis [3] In primary haemostasis, they rapidly adhere to the damaged vessel wall at the site of injury and aggregate to form a platelet plug. Platelets are known to play major effector activities in a number of additional functions, including inflammatory reactions and innate immune responses [4]. Instrumental to these activities is the ability of platelets to respond to signals from the endothelium, circulating cells, or other blood components [3]. The highly dynamic protein content of platelets has been studied using resting and activated platelets, and various subproteomes have been characterized including releasates, granules, platelet-derived microparticles (PMPs), membrane and cytoskeletal proteins, and specific PTMs occurring in platelets
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