1. Identification of inhibitors of the RGS homology domain of GRK2 by docking-based virtual screening.
- Author
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Echeverría E, Velez Rueda AJ, Cabrera M, Juritz E, Burghi V, Fabián L, Davio C, Lorenzano Menna P, and Fernández NC
- Subjects
- A549 Cells, Computer Simulation, Cyclic AMP-Dependent Protein Kinases metabolism, G-Protein-Coupled Receptor Kinase 2 chemistry, GTP-Binding Protein alpha Subunits, Gq-G11 metabolism, HeLa Cells, Humans, Mass Screening, Molecular Docking Simulation methods, Phosphorylation, Protein Binding, Protein Domains, Protein Kinase Inhibitors chemistry, Receptors, Histamine H1 metabolism, Signal Transduction, G-Protein-Coupled Receptor Kinase 2 antagonists & inhibitors, G-Protein-Coupled Receptor Kinase 2 metabolism, Protein Kinase Inhibitors pharmacology
- Abstract
Aims: G protein-coupled receptor (GPCR) kinases (GRKs) are mainly involved in the desensitization of GPCRs. Among them, GRK2 has been described to be upregulated in many pathological conditions and its crucial role in cardiac hypertrophy, hypertension, and heart failure promoted the search for pharmacological inhibitors of its activity. There have been several reports of potent and selective inhibitors of GRK2, most of them directed to the kinase domain of the protein. However, the homologous to the regulator of G protein signaling (RH) domain of GRK2 has also been shown to regulate GPCRs signaling. Herein, we searched for potential inhibitors of receptor desensitization mediated by RH domain of GRK2., Materials and Methods: We performed a docking-based virtual screening utilizing the crystal structure of GRK2 to search for potential inhibitors of the interaction between GRK2 and Gαq protein. To evaluate the biological activity of compounds we measured, calcium response of histamine H1 receptor (H1R) using Fura-2AM dye and H1R internalization by saturation binding experiments in A549 cells. GRK2(45-178)GFP translocation was determined in HeLa cells through confocal fluorescence imaging., Key Findings: We identified inhibitors of GRK2 able to reduce the RH mediated desensitization of the histamine H1 receptor and GRK2 translocation to plasma membrane. Also candidates presented adequate lipophilia and cytotoxicity profile., Significance: We obtained compounds with the ability of reducing RH mediated actions of GRK2 that can be useful as a starting point in the development of novel drug candidates aimed to treat pathologies were GRK2 plays a key role., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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