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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Functional selectivity profiling of the angiotensin II type 1 receptor using pathway-wide BRET signaling sensors

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Namkung, Yoon [1] ; LeGouill, Christian [2] ; Kumar, Sahil [1] ; Cao, Yubo [3] ; Teixeira, Larissa B. [4, 2] ; Lukasheva, Viktoriya [2] ; Giubilaro, Jenna [3] ; Simoes, Sarah C. [4] ; Longpre, Jean-Michel [5, 6] ; Devost, Dominic [3] ; Hebert, Terence E. [3] ; Pineyro, Graciela [7] ; Leduc, Richard [5, 6] ; Costa-Neto, Claudio M. [4] ; Bouvier, Michel [2] ; Laporte, Stephane A. [8, 3, 1]
Total Authors: 16
[1] McGill Univ, RI MUHC, Dept Med, Montreal, PQ H4A 3J1 - Canada
[2] Univ Montreal, IRIC, Dept Biochem & Mol Med, Montreal, PQ H3T 1J4 - Canada
[3] McGill Univ, Dept Pharmacol & Therapeut, Montreal, PQ H3G 1Y6 - Canada
[4] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, BR-14049900 Ribeirao Preto, SP - Brazil
[5] Univ Sherbrooke, Inst Pharmacol Sherbrooke, Sherbrooke, PQ J1H 5N4 - Canada
[6] Univ Sherbrooke, Dept Pharmacol Physiol, Fac Med & Hlth Sci, Sherbrooke, PQ J1H 5N4 - Canada
[7] Hop St Justine, Ctr Rech, Montreal, PQ H3T 1C5 - Canada
[8] McGill Univ, Dept Anat & Cell Biol, Montreal, PQ H3A 0C7 - Canada
Total Affiliations: 8
Document type: Journal article
Source: Science Signaling; v. 11, n. 559 DEC 4 2018.
Web of Science Citations: 7

G protein-coupled receptors (GPCRs) are important therapeutic targets that exhibit functional selectivity (biased signaling), in which different ligands or receptor variants elicit distinct downstream signaling. Understanding all the signaling events and biases that contribute to both the beneficial and adverse effects of GPCR stimulation by given ligands is important for drug discovery. Here, we report the design, validation, and use of pathway-selective bioluminescence resonance energy transfer (BRET) biosensors that monitor the engagement and activation of signaling effectors downstream of G proteins, including protein kinase C (PKC), phospholipase C (PLC), p63RhoGEF, and Rho. Combined with G protein and beta-arrestin BRET biosensors, our sensors enabled real-time monitoring of GPCR signaling at different levels in downstream pathways in both native and engineered cells. Profiling of the responses to 14 angiotensin II (AngII) type 1 receptor (AT1R) ligands enabled the clustering of compounds into different subfamilies of biased ligands and showed that, in addition to the previously reported functional selectivity between G alpha(q) and beta-arrestin, there are also biases among G protein subtypes. We also demonstrated that biases observed at the receptor and G protein levels propagated to downstream signaling pathways and that these biases could occur through the engagement of different G proteins to activate a common effector. We also used these tools to determine how naturally occurring AT1R variants affected signaling bias. This suite of BRET biosensors provides a useful resource for fingerprinting biased ligands and mutant receptors and for dissecting functional selectivity at various levels of GPCR signaling. (AU)

FAPESP's process: 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
Grantee:Claudio Miguel da Costa Neto
Support type: Research Projects - Thematic Grants