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Proteomic profiling of the thrombin-activated canine platelet secretome (CAPS)


Autoři: Signe E. Cremer aff001;  James L. Catalfamo aff002;  Robert Goggs aff003;  Stefan E. Seemann aff004;  Annemarie T. Kristensen aff001;  Marjory B. Brooks aff002
Působiště autorů: University of Copenhagen, Department of Veterinary Clinical Sciences, Copenhagen, Denmark aff001;  Cornell University, Department of Population Medicine and Diagnostic Sciences, Ithaca, New York, United States of America aff002;  Cornell University, Department Clinical Sciences, Ithaca, New York, United States of America aff003;  University of Copenhagen, Department of Veterinary and Animal Sciences, Copenhagen, Denmark aff004
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0224891

Souhrn

Domestic dogs share the same environment as humans, and they represent a valuable animal model to study naturally-occurring human disease. Platelet proteomics holds promise for the discovery of biomarkers that capture the contribution of platelets to the pathophysiology of many disease states, however, canine platelet proteomic studies are lacking. Our study objectives were to establish a protocol for proteomic identification and quantification of the thrombin-activated canine platelet secretome (CAPS), and to compare the CAPS proteins to human and murine platelet proteomic data. Washed platelets were isolated from healthy dogs, and stimulated with saline (control) or gamma-thrombin (releasate). Proteins were separated by SDS-page, trypsin-digested and analyzed by liquid chromatography and tandem mass spectrometry (MS). CAPS proteins were defined as those with a MS1-abundance ratio of two or more for releasate vs. unstimulated saline control. A total of 1,918 proteins were identified, with 908 proteins common to all dogs and 693 characterized as CAPS proteins. CAPS proteins were similar to human and murine platelet secretomes and were highly represented in hemostatic pathways. Differences unique to CAPS included replacement of platelet factor 4 with other cleavage products of platelet basic protein (e.g. interleukin-8), novel proteins (e.g. C-C motif chemokine 14), and proteins in relatively high (e.g. protease nexin-1) or low (e.g. von Willebrand factor) abundance. This study establishes the first in-depth platelet releasate proteome from healthy dogs with a reference database of 693 CAPS proteins. Similarities between CAPS and the human secretome confirm the utility of dogs as translational models of human disease, but we also identify differences unique to canine platelets. Our findings provide a resource for further investigations into disease-related CAPS profiles, and for comparative pathway analyses of platelet activation among species.

Klíčová slova:

Dogs – Fibrinogen – Chemokines – Platelet activation – Platelets – Proteomes – Proteomics – Serum proteins


Zdroje

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