title: A 2D-DIGE-based proteomic analysis reveals differences in the platelet releasate composition when comparing thrombin and collagen stimulations.
abstract: Upon stimulation, platelets release a high number of proteins (the releasate). There are clear indications that these proteins are involved in the pathogenesis of several diseases, such as atherosclerosis. In the present study we compared the platelet releasate following platelet activation with two major endogenous agonists: thrombin and collagen. Proteome analysis was based on 2D-DIGE and LC-MS/MS. Firstly, we showed the primary role of thrombin and collagen receptors in platelet secretion by these agonists; moreover, we demonstrated that GPVI is the primary responsible for collagen-induced platelet activation/aggregation. Proteomic analysis allowed the detection of 122 protein spots differentially regulated between both conditions. After excluding fibrinogen spots, down-regulated in the releasate of thrombin-activated platelets, 84 differences remained. From those, we successfully identified 42, corresponding to 37 open-reading frames. Many of the differences identified correspond to post-translational modifications, primarily, proteolysis induced by thrombin. Among others, we show vitamin K-dependent protein S, an anticoagulant plasma protein, is up-regulated in thrombin samples. Our results could have pathological implications given that platelets might be playing a differential role in various diseases and biological processes through the secretion of different subsets of granule proteins and microvesicles following a predominant activation of certain receptors.
authors: 1] Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain [2] Instituto de Investigación Sanitaria (IDIS), Santiago de Compostela, Spain [3] Departament of Pharmacology, Faculty of Pharmacy, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors: Paula
authors: P
authors: Vélez
authors: 1] Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain [2] Instituto de Investigación Sanitaria (IDIS), Santiago de Compostela, Spain [3] Departament of Pharmacology, Faculty of Pharmacy, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors: Irene
authors: I
authors: Izquierdo
authors: 1] Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain [2] Instituto de Investigación Sanitaria (IDIS), Santiago de Compostela, Spain [3] Departament of Pharmacology, Faculty of Pharmacy, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors: Isaac
authors: I
authors: Rosa
authors: 1] Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain [2] Instituto de Investigación Sanitaria (IDIS), Santiago de Compostela, Spain [3] Departament of Pharmacology, Faculty of Pharmacy, Universidade de Santiago de Compostela, Santiago de Compostela, Spain.
authors: Ángel
authors: Á
authors: García
mesh_terms: D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000494:pharmacology*; D006801:Humans; D010974:Platelet Aggregation / Q000187:drug effects; D017293:Protein S / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D011758:Pyrroles / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D053719:Tandem Mass Spectrometry; D013917:Thrombin / Q000494:pharmacology*; D057825:Two-Dimensional Difference Gel Electrophoresis; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D000906:Antibodies; C415424:N3-cyclopropyl-7-((4-(1-methylethyl)phenyl)methyl)-7H-pyrrolo(3, 2-f)quinazoline-1,3-diamine; D017293:Protein S; D020543:Proteome; D011758:Pyrroles; D011799:Quinazolines; D003094:Collagen; D013917:Thrombin
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