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Functional analysis of protein tyrosine phosphatases in thrombosis and haemostasis
Rahmouni, Souad; Hego, Alexandre; Delierneux, Céline et al.
2016In Protein Tyrosine Phosphatases: Methods and Protocols
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Abstract :
[en] Platelets are small blood cells derived from cytoplasmic fragments of megakaryocytes and play an essential role in thrombosis and haemostasis. Platelet activation depends on the rapid phosphorylation and dephosphorylation of key signaling molecules, and a number of kinases and phosphatases have been identified as major regulators of platelet function. However, the investigation of novel signaling proteins has suffered from technical limitations due to the anucleate nature of platelets and their very limited levels of mRNA and de novo protein synthesis. In the past, experimental methods were restricted to the generation of genetically modified mice and the development of specific antibodies. More recently, novel (phospho)proteomic technologies and pharmacological approaches using specific small-molecule inhibitors have added additional capabilities to investigate specific platelet proteins. In this chapter, we report methods for using genetic and pharmacological approaches to investigate the function of platelet signaling proteins. While the described experiments focus on the role of the dual-specificity phosphatase 3 (DUSP3) in platelet signaling, the presented methods are applicable to any signaling enzyme. Specifically, we describe a testing strategy that includes 1) aggregation and secretion experiments with mouse and human platelets, 2) immunoprecipitation and immunoblot assays to study platelet signaling events, 3) detailed protocols to use selected animal models in order to investigate thrombosis and haemostasis in vivo, and 4) strategies for utilizing pharmacological inhibitors on human platelets.
Disciplines :
Cardiovascular & respiratory systems
Author, co-author :
Rahmouni, Souad  ;  Université de Liège > Département des sciences cliniques > GIGA-R:Immunopath. - Maladies infect. et médec. inter. gén.
Hego, Alexandre ;  Université de Liège > Département des sciences biomédicales et précliniques > GIGA-R : Génétique humaine
Delierneux, Céline ;  Université de Liège > Département des sciences biomédicales et précliniques > GIGA-R : Génétique humaine
Wéra, Odile ;  Université de Liège > Département des sciences biomédicales et précliniques > GIGA-R : Génétique humaine
Musumeci, Lucia ;  Université de Liège > GIGA-Research
Tautz, Lutz
Oury, Cécile  ;  Université de Liège > Département des sciences biomédicales et précliniques > GIGA-R : Génétique humaine
Language :
English
Title :
Functional analysis of protein tyrosine phosphatases in thrombosis and haemostasis
Publication date :
12 August 2016
Main work title :
Protein Tyrosine Phosphatases: Methods and Protocols
Publisher :
Springer Science, New York, United States
Pages :
301-30
Peer reviewed :
Peer reviewed
Available on ORBi :
since 23 November 2015

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