Valero, TeresaDelgado-González, AntonioUnciti-Broceta, Juan DiegoCano-Cortés, VictoriaPérez-López, Ana MUnciti-Broceta, AsierSánchez Martín, Rosario M2023-01-252023-01-252018-09-04http://hdl.handle.net/10668/12851Chemical proteomics approaches are widely used to identify molecular targets of existing or novel drugs. This manuscript describes the development of a straightforward approach to conjugate azide-labeled drugs via click chemistry to alkyne-tagged cell-penetrating fluorescent nanoparticles as a novel tool to study target engagement and/or identification inside living cells. A modification of the Baeyer test for alkynes allows monitoring the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction, guaranteeing the presence of the drug on the solid support. As a proof of concept, the conjugation of the promiscuous kinase inhibitor dasatinib to Cy5-labeled nanoparticles is presented. Dasatinib-decorated fluorescent nanoparticles efficiently inhibited its protein target SRC in vitro, entered cancer cells, and colocalized with SRC in cellulo.enAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/AzidesCatalysisCell Membrane PermeabilityClick ChemistryCycloaddition ReactionDasatinibFluorescent DyesHumansNanoparticlesProteomicsresearch article30122043open access10.1021/acs.bioconjchem.8b004811520-4812https://pubs.acs.org/doi/pdf/10.1021/acs.bioconjchem.8b00481