RT Journal Article
T1 Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist
A1 Morán Lara, Luis Arturo
A1 Sowa, Marcin Andrzej
A1 Hermida Nogueira, Lidia
A1 Barrachina Núñez, María
A1 Moreira Álvarez, David
A1 Loza García, María Isabel
A1 Domínguez Medina, Eduardo
A1 García Alonso, Ángel
K1 Platelets
K1 CLEC-2
K1 High-throughput screening
K1 Small-molecule ligands
K1 Proanthocyanidin
AB AbstractBackground: 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.
PB Thieme Gruppe European Society of Cardiology
SN 0340-6245
YR 2022
FD 2022-06-28
LK https://hdl.handle.net/10347/45402
UL https://hdl.handle.net/10347/45402
LA eng
NO Morán, L. A., Di, Y., Sowa, M. A., Hermida-Nogueira, L., Barrachina, M. N., Martin, E., Clark, J. C., Mize, T. H., Eble, J. A., Moreira, D., Pollitt, A. Y., Loza, M. I., Domínguez, E., Watson, S. P., & García, Á. (2022). Katacine Is a New Ligand of CLEC-2 that Acts as a Platelet Agonist. Thrombosis and haemostasis, 122(8), 1361–1368. https://doi.org/10.1055/a-1772-1069
NO L.A.M. and M.S. are supported by the European Union’s Horizon 2020 Research and Innovation Program (Marie Sklodowska-Curie grant agreement No. 766118); S.P.W. is a British Heart Foundation Chair (CH03/003); and E.M. is supported by the Wellcome Trust (204951/Z/16/Z). A.G. is supported by the Spanish Ministry of Science and Innovation (Grant No. PID2019–108727RB-I00). J.A.E. is financially supported by the Interdisciplinary Center of Clinical Research (IZKF) of the University of Münster (grant no. Ebl-A/009/21). L.H-.N. receives financial support from the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (Centro Singular de investigación de Galicia accreditation 2019–2022, ED431G 2019/02; predoctoral grant 2018 Call).
DS Minerva
RD 27 abr 2026