Platelet membrane lipid rafts protein composition varies following GPVI and CLEC-2 receptors activation

Izquierdo, Irene; Barrachina, María N.; Hermida-Nogueira, Lidia; Casas, Vanessa; Eble, Johannes A.; Carrascal, Montserrat; Abián, Joaquín; García, Ángel

doi:10.1016/j.jprot.2019.01.014

Cited by 6 publications

(7 citation statements)

References 39 publications

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“…Western blot was performed following an established protocol 41 in an independent cohort of concentrated secretome samples collected from three donors at days 3, 7 and 21, to validate the results obtained by the proteomic SWATH-based approach. Before immunoblotting, proteins were separated in 11% SDS-PAGE gels; 10 µg of protein was loaded per lane.…”

Section: Methodsmentioning

confidence: 99%

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Hermida-Nogueira

Barrachina

Morán

et al. 2020

Sci Rep

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Leukocyte-platelet rich fibrin (L-PRF) is extensively used in the dentistry field and other clinical scenarios due to its regeneration properties. the goal of the present study was to depict the L-pRf secretome and how it changes over time. We obtained L-PRF membranes and cultured them in DMEM. The secretome was collected at days 3, 7 and 21. The secretome at day 3 was analysed by LC-MS/ MS and differences over time were analysed by Sequential Window Acquisition of all Theoretical Mass Spectra (SWATH). Overall, 705 proteins were identified in the secretome of L-PRF membranes after 3 days of culture, including growth factors (EGF, PDGFA) and proteins related to platelet and neutrophil degranulation. A total of 202 differentially secreted proteins were quantified by SWATH when comparing secretomes at days 3, 7 and 21. Most of them were enriched at day 3 such as MMP9, TSP1 and CO3. On the contrary, fibrinogen and CATS were found down-regulated at day 3. Growth factor and western blotting analysis corroborated the proteomic results. This is the most detailed proteome analysis of the L-PRF secretome to date. Proteins and growth factors identified, and their kinetics, provide novel information to further understand the wound healing properties of L-pRf. In the last two decades, the use of platelet-rich concentrates (PRC) has become very popular in different fields, especially in dentistry, due to their regeneration properties. Different scientists and companies have developed methods to obtained PRC with presence or absence of leukocytes but all of them with addition of anticoagulants to the tubes. In 2001, Choukroun et al. developed the second generation of PRC, Leukocyte platelet rich fibrin (L-PRF) 1 , which is obtained by blood centrifugation without anticoagulant in the tubes. Nowadays, L-PRF is the most commonly used PRC since it is easy to obtain and can be applied as a gel or compressed into a membrane depending on the application site. It has been mostly used in a dentistry field, especially for the treatment of osteonecrosis 2,3 , implant placement in bone-deficient sites 4 and sinus floor elevation 5. Additionally, L-PRF has started to be used in other clinical scenarios, for example for the treatment of chronic wounds, leg ulcers or diabetic foot 6 or for the treatment of ocular lesions 7. Wound healing is a complex process, which encompass different phases: haemostasis, inflammation, proliferation and remodelling. In each phase, different cell types take part in order to regenerate the tissue. Two types of blood cells, platelets and leukocytes, are among the cells that participate first. Growth factors, cytokines and other proteins released by these blood cells have chemoattractant properties that contribute to wound repair 8-10 .

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Section: Methodsmentioning

confidence: 99%

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Hermida-Nogueira

Barrachina

Morán

et al. 2020

Sci Rep

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“…Another study of the protein composition of membrane lipid rafts found large similarities after platelet stimulation via CLEC-2 or GPVI [67]. Intriguing is an observed loss of cytoskeletal proteins in the rafts from the activated platelets.…”

Section: Signalling C-type Lectin Receptor 2 (Clec-2)mentioning

confidence: 96%

“…Lipidomic mass spectrometry of complexed phosphoinositides and bound proteins showed that upon PI3K activation, many proteins can interact with 3-phosphorylated phosphoinositides through their PH domains, among which is the new adaptor protein DAPP1 [66]. Membrane lipid rafts of GPVI-stimulated platelets were also analysed in detail [67]. Oligophrenin as a small GTPase-binding protein regulator was furthermore recognised as a functional target of the GPVI-induced tyrosine kinase cascade [68].…”

Section: Collagen Receptor Glycoprotein VI (Gpvi)mentioning

confidence: 99%

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Huang

Zhang

Solari

et al. 2021

IJMS

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Platelets are small anucleate blood cells that play vital roles in haemostasis and thrombosis, besides other physiological and pathophysiological processes. These roles are tightly regulated by a complex network of signalling pathways. Mass spectrometry-based proteomic techniques are contributing not only to the identification and quantification of new platelet proteins, but also reveal post-translational modifications of these molecules, such as acetylation, glycosylation and phosphorylation. Moreover, target proteomic analysis of platelets can provide molecular biomarkers for genetic aberrations with established or non-established links to platelet dysfunctions. In this report, we review 67 reports regarding platelet proteomic analysis and signalling on a molecular base. Collectively, these provide detailed insight into the: (i) technical developments and limitations of the assessment of platelet (sub)proteomes; (ii) molecular protein changes upon ageing of platelets; (iii) complexity of platelet signalling pathways and functions in response to collagen, rhodocytin, thrombin, thromboxane A2 and ADP; (iv) proteomic effects of endothelial-derived mediators such as prostacyclin and the anti-platelet drug aspirin; and (v) molecular protein changes in platelets from patients with congenital disorders or cardiovascular disease. However, sample sizes are still low and the roles of differentially expressed proteins are often unknown. Based on the practical and technical possibilities and limitations, we provide a perspective for further improvements of the platelet proteomic field.

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“…Recently, the first proteomic comparative analysis of GEMs obtained from resting platelets and platelets activated through GPVI and CLEC-2 receptors was performed by Izquierdo et al [ 161 ]. The membrane fractions were isolated by sucrose gradient ultracentrifugation and GEMs-enriched fractions were determined by Western blot, then only the fractions enriched in flotillin-1 and lacking integrin αIIb were analyzed and GEMs proteins isolated.…”

Section: Detailed Review Of the Literature Of Proteomics Of Platelmentioning

confidence: 99%

Platelets in Healthy and Disease States: From Biomarkers Discovery to Drug Targets Identification by Proteomics

Gianazza

Brioschi

Baetta

et al. 2020

IJMS

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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.

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Platelet membrane lipid rafts protein composition varies following GPVI and CLEC-2 receptors activation

Cited by 6 publications

References 39 publications

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Deciphering the secretome of leukocyte-platelet rich fibrin: towards a better understanding of its wound healing properties

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Platelets in Healthy and Disease States: From Biomarkers Discovery to Drug Targets Identification by Proteomics

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