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Pepducin-based intervention of thrombin-receptor signaling and systemic platelet activation

Nature Medicine volume 8, pages 1161–1165 (2002)Cite this article

Abstract

Transmembrane signaling through G protein–coupled receptors (GPCRs) controls a diverse array of cellular processes including metabolism, growth, motility, adhesion, neuronal signaling and blood coagulation. The numerous GPCRs and their key roles in both normal physiology and disease have made them the target for more than 50% of all prescribed drugs. GPCR agonists and antagonists act on the extracellular side of the receptors, whereas the intracellular surface has not yet been exploited for development of new therapeutic agents. Here, we demonstrate the utility of novel cell-penetrating peptides, termed 'pepducins', that act as intracellular inhibitors of signal transference from receptors to G proteins1. Attachment of a palmitate lipid to peptides based on the third intracellular loop of protease-activated receptor 1 (PAR1)2 or PAR4 (refs. 3–5) yielded potent inhibitors of thrombin-mediated aggregation of human platelets. Infusion of the anti-PAR4 pepducin into mice extended bleeding time and protected against systemic platelet activation, consistent with the phenotype of PAR4-deficient mice. We show that pepducins might be used to ascertain the physiological roles of GPCRs and rapidly determine the potential therapeutic value of blockade of a particular signaling pathway.

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Figure 1: Palmitoylated peptides derived from the i3 loop of PAR1 and PAR4 act as antagonists of receptor–G-protein signaling in human platelets.
Figure 2: The human PAR4 pepducin, P4pal-10, inhibits aggregation of human and murine platelets by thrombin.
Figure 3: The human P4pal-10 pepducin prolongs bleeding time and protects against systemic platelet activation in mice.

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Acknowledgements

Acknowledgments This work was supported by a Natalie V. Zucker Award (to L.C.) and National Institutes of Health grants R01HL64701, R01HL57905 (to A.K.).

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  1. Division of Hematology/Oncology, New England Medical Center and Departments of Medicine and Biochemistry, Molecular Cardiology Research Institute, Tufts University School of Medicine, Boston, Massachusetts, USA

    Lidija Covic, Meghna Misra, Jehangir Badar, Christopher Singh & Athan Kuliopulos

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  1. Lidija Covic
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  2. Meghna Misra
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Correspondence to Athan Kuliopulos.

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Tufts-New England Medical Center Hospital has submitted a patent application covering pepducins.

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Covic, L., Misra, M., Badar, J. et al. Pepducin-based intervention of thrombin-receptor signaling and systemic platelet activation. Nat Med 8, 1161–1165 (2002). https://doi.org/10.1038/nm760

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