A novel di‐peptide Met‐Glu from collagen hydrolysates inhibits platelet aggregation and thrombus formation via regulation of Gq‐mediated signaling

Abstract

Increasing evidence has shown that collagen peptides had various biological activities. In this study, a novel antiplatelet peptide Met‐Glu (ME) was separated and identified from silver carp skin by YMC ODS‐A C18 separation and ESI‐MS/MS analysis. Peptide ME inhibited platelet aggregation and secretion of platelet granules induced by ADP, thrombin and collagen, and significantly attenuated ferric chloride‐induced thrombus formation in rats. It did not prolong the bleeding time in mice even at the dose of 300 μmol/kg body weight that showed potent anti‐thrombosis effects. Additionally, peptide ME targeted at Gq‐protein to downregulate the phosphorylation of PLCβ, an important upstream effector of PI3K/Akt and Erk/MAPK signaling to inhibit intracellular calcium ion mobilization. These results suggest that peptide ME inhibited thrombosis in vivo and inhibited Gq‐mediated signaling in platelets, indicating the possibility that ME could potentially be developed as a novel therapeutic agent in the prevention and treatment of thrombotic diseases. Practical applications Cardiovascular diseases (CVDs) are the leading cause of mortality and morbidity worldwide. The proximal cause of CVDs is intravascular thrombosis formation, which mostly results from platelet activation, aggregation, and granules secretion. Traditional drugs in the prevention of thrombotic disease, such as aspirin and clopidogrel, are still limited for their side effects, especially bleeding complications. Collagen is a natural source for bioactive peptides and our previous study has shown that collagen peptides could inhibit platelet aggregation in vitro. Understanding the mechanism of collagen peptides on regulation of platelet activation and their in vivo anti‐thrombosis activities were important for the development of novel‐specific medical food in the prevention of thrombotic diseases.

A Collagen‐Derived Oligopeptide from Salmo salar Collagen Hydrolysates Restrains Atherogenesis in ApoE–/– Mice via Targeting P2Y12 Receptor

“…Western Blotting: Immunoblot assays were performed as described previously. [36] Aliquots of platelets were preincubated with vehicle, peptide OG-5 or specific inhibitors, and then stimulated with or without agonist. The reaction was stopped by the addition of RIPA buffer containing a protease inhibitor PMSF.…”

Review on Extraction, Modification, and Synthesis of Natural Peptides and Their Beneficial Effects on Skin

“…This was evident in peptides isolated from fermented milk (yogurt) [ 28 ] and rapeseed (Brassica campestris L.) [ 29 ] by preparative reversed-phase HPLC, then purified and analyzed by ESI-MS/MS. Notably, a novel peptide with antiplatelet activity was isolated and identified from silver carp ( Hypophthalmichthys molitrix ) skin by the same method [ 30 ], as was a novel peptide purified by solid-phase extraction and HPLC from the interleaf of banana ( Musa paradisiaca ) plants, with its primary structure determined by MS and amino acid analysis [ 31 ].…”

Novel Peptide Motifs Containing Asp-Glu-Gly Target P2Y12 and Thromboxane A2 Receptors to Inhibit Platelet Aggregation and Thrombus Formation

“…Immunoblot assays were performed as described previously. 22 Aliquots of washed platelets were preincubated with vehicle or peptide DEGP and then stimulated with or without the agonist U46619. PRP was used when stimulated with 2MeS-ADP.…”