PXD070478

PXD070478 is an original dataset announced via ProteomeXchange.

Title	Mapping the protein-protein interaction interface between recombinant CB1R and GlyT2 using cross-linking mass spectrometry
Description	This project aims to elucidate a novel molecular mechanism driving mechanical allodynia, a debilitating neuropathic pain condition. We focus on a spinal cord “allodynia gate” circuit in which nerve injury leads to disinhibition of protein kinase Cγ (PKCγ) excitatory neurons. Our findings reveal that this disinhibition is mediated by endocannabinoid (eCB) signaling: eCBs released from PKCγ neurons activate presynaptic cannabinoid receptor 1 (CB1R) located on glycinergic (Gly) neurons. A key event in this process is the functional interaction between CB1R and glycine transporter 2 (GlyT2) within Gly neurons. However, the structural basis of the CB1R–GlyT2 complex remained poorly understood. The mass spectrometry data presented in this repository directly address this knowledge gap. Using chemical cross-linking mass spectrometry (CX-MS) with the BS³ cross-linker, we mapped specific interaction interfaces between the N-terminal domain of CB1R and defined extracellular and intracellular domains of GlyT2. The protein–protein interaction identified by CX-MS was independently validated by co-immunoprecipitation (Co-IP) assays. The structural insights derived from this CX-MS dataset enabled the rational design of a disrupting peptide, Tat-STVKIAK-KFERQ, which targets the CB1R–GlyT2 interface. This peptide effectively prevented and alleviated nerve injury–induced mechanical allodynia in mice. Thus, the data provided here not only provide critical structural evidence for a pivotal protein complex in neuropathic pain but also establish a foundation for a targeted therapeutic strategy.
HostingRepository	PRIDE
AnnounceDate	2026-02-13
AnnouncementXML	Submission_2026-02-13_04:49:00.353.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Qun Wang
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-11-07 14:55:48	ID requested
⏵ 1	2026-02-13 04:49:01	announced

10.1016/j.neuron.2025.11.016;

Wang Q, Jiang Z, Huang C, Zhang X, Zhao J, Ma H, Qi J, Li J, Yang Y, He X, Du S, Li Y, Fang Z, Liu P, Zuo W, Liu P, Wang Y, Zhao G, Gu N, Xiong L, Hou W, Dong H, Lu Y, R and GlyT2 interaction in spinal glycinergic circuits drives neuropathic mechanical pain. Neuron, 114(5):884-902.e12(2026) [pubmed]

submitter keyword: CB1R protein, CX-MS, GlyT2 protein

Yan Lu
contact affiliation	Department of Anesthesiology and Perioperative Medicine, Xijing Hospital,the Fourth Military Medical University, Xi'an 710032, China
contact email	yanlu001@fmmu.edu.cn
lab head
Qun Wang
contact affiliation	Xijing Hospital, fourth military medical university
contact email	wqfmmu77@163.com
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/02/PXD070478

PRIDE project URI

• PRIDE
  1. PXD070478
     1. Label: PRIDE project
     2. Name: Mapping the protein-protein interaction interface between recombinant CB1R and GlyT2 using cross-linking mass spectrometry