A peptide from the staphylococcal protein Efb binds P-selectin and inhibits the interaction of platelets with leukocytes

Stuart Wallis, Nina Wolska, Hanna Englert, Mareike Posner, Abhishek Upadhyay, Thomas Renne, Ian Eggleston, Stefan Bagby, Giordano Pula

Research output: Contribution to journalArticlepeer-review

2 Citations (SciVal)
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Abstract

Aims
P-selectin is a key surface adhesion molecule for the interaction of platelets with leukocytes. We have shown previously that the N-terminal domain of S. aureus extracellular fibrinogen-binding protein (Efb) binds to P-selectin and interferes with platelet-leukocyte aggregate formation. Here, we aimed to identify the minimal Efb motif required for binding platelets and to characterise its ability to interfering with the formation of platelet-leukocyte aggregates.

Methods and Results
Using a library of synthetic peptides, we mapped the platelet-binding site to a continuous 20 amino acid stretch. The peptide Efb68-87 was able to bind to resting and, to a greater extent, thrombin-stimulated platelets in the absence of fibrinogen. Dot blots, pull-down assays and P-selectin glycoprotein ligand-1 (PSGL-1) competitive binding experiments identified P-selectin as the cellular docking site mediating Efb68-87 platelet binding. Accordingly, Efb68-87 did not bind to other blood cells and captured platelets from human whole blood under low shear stress conditions. Efb68-87 did not affect platelet activation as tested by aggregometry, flow cytometry and immunoblotting, but inhibited the formation of platelet-leukocyte aggregates (PLAs). Efb68-87 also interfered with the platelet-dependent stimulation of neutrophil extracellular traps (NETs) formation in vitro.

Conclusions
We have identified Efb68-87 as a novel selective platelet-binding peptide. Efb68-87 binds directly to P-selectin and inhibits interactions of platelets with leukocytes that lead to PLA and NET formation. As PLAs and NETs play a key role in thromboinflammation, Efb68-87 is an exciting candidate for the development of novel selective inhibitors of the proinflammatory activity of platelets.
Original languageEnglish
Pages (from-to)729-741
Number of pages13
JournalJournal of Thrombosis and Haemostasis
Volume20
Issue number3
Early online date30 Nov 2021
DOIs
Publication statusPublished - 31 Mar 2022

Keywords

  • leukocyte
  • neutrophil extracellular trap
  • P-selectin
  • platelet
  • platelet-leukocyte aggregate
  • thromboinflammation

ASJC Scopus subject areas

  • Hematology

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