Human -Synuclein Inhibits Platelets Aggregation in vitro by Interfering with the -Thrombin/Protease-Activated Receptor 1 Functional Axis

Pontarollo, G.; Acquasaliente, L.; Radu, C. M.; Peterle, D.; Artusi, I.; Pagotto, A.; Uliana, F.; Simioni, P.; Negro, A.; De Filippis, V.

doi:10.1101/2021.03.28.437436

bioRxiv · 10.1101/2021.03.28.437436

Human -Synuclein Inhibits Platelets Aggregation in vitro by Interfering with the -Thrombin/Protease-Activated Receptor 1 Functional Axis

Pontarollo, G.·Acquasaliente, L.·Radu, C. M.·Peterle, D.·Artusi, I.·Pagotto, A.·Uliana, F.·Simioni, P.·Negro, A.·De Filippis, V.

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Abstract

-Synuclein (Syn) is a small (140 amino acids) disordered, acidic (pI: 4.7) protein, highly conserved in vertebrates and implicated in the pathogenesis of Parkinsons disease (PD), a neurodegenerative disease characterized by the deposition of Syn amyloid fibrils in dopaminergic neurons. Beyond the central nervous system, significant expression of Syn has also been measured in the blood (~1 M), where platelets are the main cellular hosts of Syn. Although the pathological implication of Syn in PD is widely accepted, the physiological role of blood Syn is still elusive. Starting from the notion that platelets are either the major cellular reservoir of Syn in the blood and, concomitantly, act as key players in hemostasis, being activated also by -thrombin (T) via cleavage of protease-activated receptors (PARs), we decided to investigate the possibility that Syn could modulate platelet activation by interfering with the T-PAR functional axis. Using multiple electrode aggregometry, i.e. a fast and specific platelet-function-testing method, as well as steady-state fluorescence spectroscopy, surface plasmon resonance, and fluorescence microscopy, we show here that monomeric Syn functions as a negative regulator of T-mediated platelets activation. Syn acts either directly, via competitive inhibition of PAR1 activation by T and TRAP6 agonist, and indirectly, by scavenging T on the platelet plasma membrane. A simple electrostatic model of Syn platelet antiaggregating effect is proposed and the possible role of the protein at the interplay of amyloidosis and thrombosis is discussed.

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Pontarollo, G., Acquasaliente, L., Radu, C. M., Peterle, D., Artusi, I., Pagotto, A., Uliana, F., Simioni, P., Negro, A., De Filippis, V.. 2021-03-29. Human -Synuclein Inhibits Platelets Aggregation in vitro by Interfering with the -Thrombin/Protease-Activated Receptor 1 Functional Axis. https://doi.org/10.1101/2021.03.28.437436

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