1. Discovery of a dual Ras and ARF6 inhibitor from a GPCR endocytosis screen
- Author
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Jean Duchaine, Stéphane A. Laporte, Etienne Khoury, Alexandre Beautrait, Yoon Namkung, Michel Bouvier, Mónica Lara-Márquez, Tomasz Maciej Stepniewski, Doris A. Schuetz, Audrey Claing, Olivier Radresa, Sylvain Armando, Anne Marinier, Nathalie Lamarche-Vane, Jana Selent, Jenna Giubilaro, and Shirley Campbell
- Subjects
MAPK/ERK pathway ,Bioluminescence Resonance Energy Transfer Techniques ,Science ,media_common.quotation_subject ,General Physics and Astronomy ,Small G Protein ,Hormone receptors ,Molecular Dynamics Simulation ,Endocytosis ,General Biochemistry, Genetics and Molecular Biology ,Article ,Receptors, G-Protein-Coupled ,Cell Line, Tumor ,Drug Discovery ,Humans ,Enzyme Inhibitors ,Receptor ,Internalization ,Protein kinase B ,G protein-coupled receptor ,media_common ,Cell Proliferation ,Multidisciplinary ,Binding Sites ,Chemistry ,ADP-Ribosylation Factors ,Receptor Protein-Tyrosine Kinases ,General Chemistry ,Angiotensin II ,Cell biology ,HEK293 Cells ,ADP-Ribosylation Factor 6 ,Screening ,ras Proteins ,Signal Transduction - Abstract
Internalization and intracellular trafficking of G protein-coupled receptors (GPCRs) play pivotal roles in cell responsiveness. Dysregulation in receptor trafficking can lead to aberrant signaling and cell behavior. Here, using an endosomal BRET-based assay in a high-throughput screen with the prototypical GPCR angiotensin II type 1 receptor (AT1R), we sought to identify receptor trafficking inhibitors from a library of ~115,000 small molecules. We identified a novel dual Ras and ARF6 inhibitor, which we named Rasarfin, that blocks agonist-mediated internalization of AT1R and other GPCRs. Rasarfin also potently inhibits agonist-induced ERK1/2 signaling by GPCRs, and MAPK and Akt signaling by EGFR, as well as prevents cancer cell proliferation. In silico modeling and in vitro studies reveal a unique binding modality of Rasarfin within the SOS-binding domain of Ras. Our findings unveil a class of dual small G protein inhibitors for receptor trafficking and signaling, useful for the inhibition of oncogenic cellular responses., While Ras is a promising target for cancer therapy, development of inhibitors targeting Ras signaling has proven challenging. Here, the authors report the discovery of Rasarfin, a small molecule from a phenotypic screen on G protein-coupled receptor (GPCR) endocytosis that acts as a dual Ras and ARF6 inhibitor.
- Published
- 2021