Experimental validation of computerised models of clustering of platelet glycoprotein receptors that signal via tandem SH2 domain proteins.

Autor: Maqsood Z; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Alacris Theranostics, GmbH, Berlin, Germany.; Rudolf Virchow Center for Integrative and Translation Bioimaging, University of Würzburg and Institute of Experimental Biomedicine I, University Hospital of Würzburg, Würzburg, Germany., Clark JC; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom., Martin EM; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Cheung YFH; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Leibniz-Institut für Analytische Wissenschaften-ISAS-e. V., Dortmund, Germany.; School of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, The Netherlands., Morán LA; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Watson SET; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Pike JA; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom., Di Y; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Poulter NS; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom., Slater A; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Lange BMH; Alacris Theranostics, GmbH, Berlin, Germany., Nieswandt B; Rudolf Virchow Center for Integrative and Translation Bioimaging, University of Würzburg and Institute of Experimental Biomedicine I, University Hospital of Würzburg, Würzburg, Germany., Eble JA; Institute of Physiological Chemistry and Pathobiochemistry, University of Münster, Münster, Germany., Tomlinson MG; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom.; Department of Biosciences, University of Birmingham, Birmingham, United Kingdom., Owen DM; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom.; Institute of Immunology and Immunotherapy, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom., Stegner D; Rudolf Virchow Center for Integrative and Translation Bioimaging, University of Würzburg and Institute of Experimental Biomedicine I, University Hospital of Würzburg, Würzburg, Germany., Bridge LJ; Faculty of Environment & Technology, Department of Computer Science and Creative Technologies, University of the West England, Bristol, United Kingdom., Wierling C; Alacris Theranostics, GmbH, Berlin, Germany., Watson SP; Institute of Cardiovascular Sciences, IBR Building, College of Medical and Dental Sciences, University of Birmingham, Birmingham, United Kingdom.; Centre of Membrane Proteins and Receptors (COMPARE), Universities of Birmingham and Nottingham, Birmingham, United Kingdom.; School of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, Maastricht, The Netherlands.
Jazyk: angličtina
Zdroj: PLoS computational biology [PLoS Comput Biol] 2022 Nov 28; Vol. 18 (11), pp. e1010708. Date of Electronic Publication: 2022 Nov 28 (Print Publication: 2022).
DOI: 10.1371/journal.pcbi.1010708
Abstrakt: The clustering of platelet glycoprotein receptors with cytosolic YxxL and YxxM motifs, including GPVI, CLEC-2 and PEAR1, triggers activation via phosphorylation of the conserved tyrosine residues and recruitment of the tandem SH2 (Src homology 2) domain effector proteins, Syk and PI 3-kinase. We have modelled the clustering of these receptors with monovalent, divalent and tetravalent soluble ligands and with transmembrane ligands based on the law of mass action using ordinary differential equations and agent-based modelling. The models were experimentally evaluated in platelets and transfected cell lines using monovalent and multivalent ligands, including novel nanobody-based divalent and tetravalent ligands, by fluorescence correlation spectroscopy. Ligand valency, receptor number, receptor dimerisation, receptor phosphorylation and a cytosolic tandem SH2 domain protein act in synergy to drive receptor clustering. Threshold concentrations of a CLEC-2-blocking antibody and Syk inhibitor act in synergy to block platelet aggregation. This offers a strategy for countering the effect of avidity of multivalent ligands and in limiting off-target effects.
Competing Interests: We have read the journal’s policy and the authors of this manuscript have the following competing interests: SPW and NSP declare a patent to protect the nanobodies which was filed on 21st December 2021 (PCT/EP2021/087129: ‘Nanobody’). All other authors have declared that they have no conflict of interest.
(Copyright: © 2022 Maqsood et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje