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Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist
Background CLEC-2 is a platelet receptor with an important role in thromboinflammation but a minor role in hemostasis. Two endogenous ligands of CLEC-2 have been identified, the transmembrane protein podoplanin and iron-containing porphyrin hemin, which is formed following hemolysis from red blood...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Georg Thieme Verlag KG
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393086/ https://www.ncbi.nlm.nih.gov/pubmed/35170009 http://dx.doi.org/10.1055/a-1772-1069 |
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author | Morán, Luis A. Di, Ying Sowa, Marcin A. Hermida-Nogueira, Lidia Barrachina, María N. Martin, Eleyna Clark, Joanne C. Mize, Todd H. Eble, Johannes A. Moreira, David Pollitt, Alice Y. Loza, María I. Domínguez, Eduardo Watson, Steve P. García, Ángel |
author_facet | Morán, Luis A. Di, Ying Sowa, Marcin A. Hermida-Nogueira, Lidia Barrachina, María N. Martin, Eleyna Clark, Joanne C. Mize, Todd H. Eble, Johannes A. Moreira, David Pollitt, Alice Y. Loza, María I. Domínguez, Eduardo Watson, Steve P. García, Ángel |
author_sort | Morán, Luis A. |
collection | PubMed |
description | Background CLEC-2 is a platelet receptor with an important role in thromboinflammation but a minor role in hemostasis. Two endogenous ligands of CLEC-2 have been identified, the transmembrane protein podoplanin and iron-containing porphyrin hemin, which is formed following hemolysis from red blood cells. Other exogenous ligands such as rhodocytin have contributed to our understanding of the role of CLEC-2. Objectives To identify novel CLEC-2 small-molecule ligands to aid therapeutic targeting of CLEC-2. Methods ALPHA screen technology has been used for the development of a high-throughput screening (HTS) assay recapitulating the podoplanin–CLEC-2 interaction. Light transmission aggregometry was used to evaluate platelet aggregation. Immunoprecipitation and western blot were used to evaluate direct phosphorylation of CLEC-2 and downstream protein phosphorylation. Autodock vina software was used to predict the molecular binding site of katacine and mass spectrometry to determine the polymeric nature of the ligand. Results and Conclusion We developed a CLEC-2–podoplanin interaction assay in a HTS format and screened 5,016 compounds from a European Union-open screen library. We identified katacine, a mixture of polymers of proanthocyanidins, as a novel ligand for CLEC-2 and showed that it induces platelet aggregation and CLEC-2 phosphorylation via Syk and Src kinases. Platelet aggregation induced by katacine is inhibited by the anti-CLEC-2 monoclonal antibody fragment AYP1 F(ab)′2. Katacine is a novel nonprotein ligand of CLEC-2 that could contribute to a better understanding of CLEC-2 activation in human platelets. |
format | Online Article Text |
id | pubmed-9393086 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Georg Thieme Verlag KG |
record_format | MEDLINE/PubMed |
spelling | pubmed-93930862022-08-22 Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist Morán, Luis A. Di, Ying Sowa, Marcin A. Hermida-Nogueira, Lidia Barrachina, María N. Martin, Eleyna Clark, Joanne C. Mize, Todd H. Eble, Johannes A. Moreira, David Pollitt, Alice Y. Loza, María I. Domínguez, Eduardo Watson, Steve P. García, Ángel Thromb Haemost Background CLEC-2 is a platelet receptor with an important role in thromboinflammation but a minor role in hemostasis. Two endogenous ligands of CLEC-2 have been identified, the transmembrane protein podoplanin and iron-containing porphyrin hemin, which is formed following hemolysis from red blood cells. Other exogenous ligands such as rhodocytin have contributed to our understanding of the role of CLEC-2. Objectives To identify novel CLEC-2 small-molecule ligands to aid therapeutic targeting of CLEC-2. Methods ALPHA screen technology has been used for the development of a high-throughput screening (HTS) assay recapitulating the podoplanin–CLEC-2 interaction. Light transmission aggregometry was used to evaluate platelet aggregation. Immunoprecipitation and western blot were used to evaluate direct phosphorylation of CLEC-2 and downstream protein phosphorylation. Autodock vina software was used to predict the molecular binding site of katacine and mass spectrometry to determine the polymeric nature of the ligand. Results and Conclusion We developed a CLEC-2–podoplanin interaction assay in a HTS format and screened 5,016 compounds from a European Union-open screen library. We identified katacine, a mixture of polymers of proanthocyanidins, as a novel ligand for CLEC-2 and showed that it induces platelet aggregation and CLEC-2 phosphorylation via Syk and Src kinases. Platelet aggregation induced by katacine is inhibited by the anti-CLEC-2 monoclonal antibody fragment AYP1 F(ab)′2. Katacine is a novel nonprotein ligand of CLEC-2 that could contribute to a better understanding of CLEC-2 activation in human platelets. Georg Thieme Verlag KG 2022-06-28 /pmc/articles/PMC9393086/ /pubmed/35170009 http://dx.doi.org/10.1055/a-1772-1069 Text en The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. ( https://creativecommons.org/licenses/by-nc-nd/4.0/ ) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License, which permits unrestricted reproduction and distribution, for non-commercial purposes only; and use and reproduction, but not distribution, of adapted material for non-commercial purposes only, provided the original work is properly cited. |
spellingShingle | Morán, Luis A. Di, Ying Sowa, Marcin A. Hermida-Nogueira, Lidia Barrachina, María N. Martin, Eleyna Clark, Joanne C. Mize, Todd H. Eble, Johannes A. Moreira, David Pollitt, Alice Y. Loza, María I. Domínguez, Eduardo Watson, Steve P. García, Ángel Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title | Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title_full | Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title_fullStr | Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title_full_unstemmed | Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title_short | Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist |
title_sort | katacine is a new ligand of clec-2 that acts as a platelet agonist |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9393086/ https://www.ncbi.nlm.nih.gov/pubmed/35170009 http://dx.doi.org/10.1055/a-1772-1069 |
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