PXD047827-1

PXD047827 is an original dataset announced via ProteomeXchange.

Title	Investigation of GPCR-G protein coupling by Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) : the GPBAR1 case.
Description	G protein-coupled receptors (GPCRs) are key classes of proteins for drug discovery research as many of them are still orphans and their function poorly known. Their activation mechanism is roughly divided in two states : active and unactive states however related dynamics to switch from one state to another are poorly understood. The rise of cryogenic electron microscopy (cryo-EM) allowed to determine numerous GPCR high resolution structures in complex with G-proteins or not during the last couple of years. In this structural biology landscape, a few alternative techniques such as Hydrogen-Deuterium eXchange coupled to Mass Spectrometry (HDX-MS) have emerged as particularly suitable to characterize protein dynamics including membrane proteins. Here, for the first time, we optimized an HDX-MS workflow to characterize a class A GPCR, the G-protein-coupled bile acid receptor (also known as Gpbar1 or TGR5), in complex with G-proteins and a stabilizing nanobody. We, first, extensively describe the HDX-MS method optimization and related useful information we got regarding folding of the five proteins mix. Second, we deciphered the dynamics of each protein partner in the apo and holo states from HDX labelling experiments and performed data interpretation integrating dynamics information from HDX-MS and high-resolution structure from cryo-EM. This work highlights the complementary of both techniques for deep GPCR-Protein G coupling understanding in a drug discovery context.
HostingRepository	PRIDE
AnnounceDate	2025-09-08
AnnouncementXML	Submission_2025-09-07_17:41:23.585.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jérôme CASTEL
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Xevo G2-XS QTof

Revision	Datetime	Status	ChangeLog Entry
0	2023-12-14 04:45:21	ID requested
⏵ 1	2025-09-07 17:41:24	announced

10.1038/s41598-025-16529-w;

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(1):31517(2025) [pubmed]

submitter keyword: cryo-EM,GPBAR1, G protein, GPCR-G protein, TGR5, HDX-MS, GPCR

Thomas BOTZANOWSKI, HDX-MS lead
contact affiliation	NovAliX, Department of Biophysics, 16 Rue d’Ankara, 67000, Strasbourg, France; orcid.org/0000-0001-6229-4524
contact email	thomas.botzanowski.pro@gmail.com
lab head
Jérôme CASTEL
contact affiliation	Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO), Institut Pluridisciplinaire Hubert Curien – IPHC. CNRS UMR 7178, Université de Strasbourg, France NovAliX, Department of Biophysics, 16 Rue d’Ankara, 67000, Strasbourg, France
contact email	jerome.castel@etu.unistra.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD047827
     1. Label: PRIDE project
     2. Name: Investigation of GPCR-G protein coupling by Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) : the GPBAR1 case.