9GYO

CryoEM structure of Gs-coupled GPBAR with small molecule agonist P395

Summary for 9GYO

• Entry DOI: 10.2210/pdb9gyo/pdb
• EMDB information: 51700
• Descriptor: Isoform Gnas-2 of Guanine nucleotide-binding protein G(s) subunit alpha isoforms short, Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1, Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2, ... (7 entities in total)
• Functional Keywords: g-protein coupled receptor, transmembrane protein, g protein signaling, bile acid-binding., membrane protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 5
• Total formula weight: 159202.83

Authors

Frechard, A.,Brooks, I. (deposition date: 2024-10-02, release date: 2024-10-09, Last modification date: 2025-09-17)

Primary citation

Castel, J.,Botzanowski, T.,Brooks, I.,Frechard, A.,Bey, G.,Schroeter, M.,Del Nero, E.,Debaene, F.,Ciesielski, F.,Zeyer, D.,Cianferani, S.,Morales, R.
Deciphering molecular determinants of GPBAR1-Gs protein interactions by HDX-MS and cryo-EM.
Sci Rep, 15:31517-31517, 2025

PubMed Abstract: Many physiological processes are dependent on G protein-coupled receptors (GPCRs), the biggest family of human membrane proteins and a significant class of therapeutic targets. Once activated by external stimuli, GPCRs use G proteins and arrestins as transducers to generate second messengers and trigger downstream signaling, leading to diverse signaling profiles. The G protein-coupled bile acid receptor 1 (GPBAR1, also known as Takeda G protein-coupled receptor 5, TGR5) is a class A bile acid membrane receptor that regulates energy homeostasis and glucose and lipid metabolism. GPBAR1/Gs protein interactions are implicated in the prevention of diabetes and the reduction of inflammatory responses, making GPBAR1 a potential therapeutic target for metabolic disorders. Here, we present combined hydrogen/deuterium exchange mass spectrometry (HDX-MS) and cryo-electron microscopy (cryo-EM) to identify the molecular determinants of GPBAR1 conformational dynamics upon G protein binding. Thanks to extensive optimization, we achieved over 75% sequence coverage by HDX-MS of a complete GPCR complex and a 2.5 Å resolution structure by cryo-EM, both of which are state-of-the-art. Altogether, our results provide information on the under-investigated GPBAR1 binding mode to its cognate G protein, pinpointing the synergic and powerful combination of higher cryo-EM and lower HDX-MS resolution techniques to dissect GPCR/G protein binding characteristics.

PubMed: 40858717
DOI: 10.1038/s41598-025-16529-w

Experimental method

ELECTRON MICROSCOPY (2.5 Å)