Dataset Information

Mapping the protein-protein interaction interface between recombinant CB1R and GlyT2 using cross-linking mass spectrometry

ABSTRACT: 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.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Qun Wang

LAB HEAD: Yan Lu

PROVIDER: PXD070478 | Pride | 2026-02-13

REPOSITORIES: Pride

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Publications

CB<sub>1</sub>R and GlyT2 interaction in spinal glycinergic circuits drives neuropathic mechanical pain.

Wang Qun Q Jiang Zhenhua Z Huang Chenchen C Zhang Xiao X Zhao Jianshuai J Ma Hongwei H Qi Jing J Li Jin J Yang Yongheng Y He Xiaolan X Du Shibin S Li You Y Fang Zongping Z Liu Peng P Zuo Wenqiang W Liu Peizheng P Wang Ye Y Zhao Guangchao G Gu Nan N Xiong Lize L Hou Wugang W Dong Hailong H Lu Yan Y

Neuron 20260130

Mechanical allodynia, a manifestation of neuropathic pain, affects patients' well-being. Our previous research identified a spinal "allodynia gate" involving glycinergic (Gly) inhibitory and protein kinase Cγ (PKCγ) excitatory neurons. Nerve injury disrupts this circuit, leading to allodynia. However, the disinhibition mechanisms remain unclear. Here, genetically modified mouse lines were used to demonstrate that nerve injury triggers endocannabinoid (eCB) release from PKCγ neurons, activating c ...[more]

PMID: 41619725

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			Action	DRS
	CNR1-SLC6A5_extra_XLpeptides.csv	Csv
	CNR1-SLC6A5_intra_XLpeptides.csv	Csv
	CSZ-extra.mgf	Mgf
	CSZ-extra.raw	Raw
	CSZ_intra.mgf	Mgf