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Space-time characterization of G protein-coupled receptors (GPCRs) signal transduction through new bioluminescence resonance energy transfer (BRET)-based biosensors

Grant number: 18/13655-9
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): April 01, 2019
Effective date (End): March 06, 2022
Field of knowledge:Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
Principal Investigator:Rita de Cassia Aleixo Tostes Passaglia
Grantee:Diego Ângelo Duarte
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:12/20148-0 - Development of new ligands/drugs with selective agonism action (biased agonism) for receptors of the renin-angiotensin and kallikrein-kinin systems: new properties and new biotechnological applications, AP.TEM

Abstract

The G protein-coupled receptor (GPCR) superfamily mediates the majority of the physiological processes and is target for approximately 40% of the drugs currently in the market. Despite the recent advances in the study of GPCR signaling, the role of lipid rafts in GPCRs signaling regulation is still not defined, probably due to the lack of accurate enough technologies to study lipid rafts in living cells. The aim of this project is to investigate how lipid rafts can regulate the GPCRs signal transduction and to study the involvement of receptor interaction to ²-arrestin or different classes of G protein in this regulation. For that, we will analyse the location and membrane translocation of different GPCRs and effectors (e.g. G proteins and ²-arrestins) using biosensors based on bioluminescence resonance energy transfer (BRET) technology that specifically localize to lipid rafts or non-rafts domains in the plasma membrane. Such biosensors were developed and characterized during the BEPE training period carried on during my PhD in the renowned laboratory headed by Professor Michel Bouvier at the University of Montreal, Canada. We will also analyze the modulation of signaling from the different compartments on the cell surface, and receptors and effectors location will also be analyzed by super-resolution confocal microscopy. We believe that the elucidation of the of lipid rafts role in the regulation of GPCR mediated signaling and the molecular mechanisms involved in this regulation will generate a sound contribution to the GPCR field, and shall as well contribute for a better understanding of mechanisms of action and development of novel drugs.

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DUARTE, DIEGO A.; PARREIRAS-E-SILVA, LUCAS T.; OLIVEIRA, EDUARDO B.; BOUVIER, MICHEL; COSTA-NETO, CLAUDIO M.. Angiotensin II Type 1 Receptor Tachyphylaxis Is Defined by Agonist Residence Time. Hypertension, v. 79, n. 1, p. 115-125, . (16/15914-6, 18/13655-9, 12/20148-0, 16/24120-3, 14/09893-0)

Please report errors in scientific publications list by writing to: cdi@fapesp.br.