PXD068157-1

PXD068157 is an original dataset announced via ProteomeXchange.

Title	Proteomic identification of stomatin-associated proteins in HEK293T cells by co-immunoprecipitation and APEX2-based proximity labeling mass spectrometry
Description	To identify proteins enriched in functional membrane microdomains organized by stomatin—hereafter referred to as STOM cargo proteins—we designed mass spectrometry (MS)–based approaches combining co-immunoprecipitation (co-IP) MS and enzymatic proximity labeling MS. For these experiments, we generated a series of stable HEK293T cell lines via lentiviral transduction. For the Co-IP MS analysis, we employed GFP-STOM-WT to capture stomatin and its interacting partners from cellular lysates. Additionally, we generated a cytoplasm-localized GFP-NES control by fusing GFP to a nuclear export signal (NES), enabling subtraction of GFP tag-derived contaminants. Enrichment was performed using a GFP nanobody. GDN was chosen for membrane extraction due to its ability to preserve the membrane region within the stomatin assemblies. This approach enriches cargo proteins interacting with stomatin directly or embedded in the membrane region within the stomatin bowl-like structure. We also applied proximity labeling MS to profile STOM cargo proteins. The labeling enzyme was fused at the N-terminus of stomatin, such that labeled proteins should reside outside the closed stomatin bowl.
HostingRepository	iProX
AnnounceDate	2025-09-06
AnnouncementXML	Submission_2026-09-09_19:11:20.311.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lu Yan
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-07 23:52:26	ID requested
⏵ 1	2026-09-09 19:11:20	announced

Dataset with its publication pending

submitter keyword: stomatin, human, HEK293T,co-IP, AP-MS, APEX2

Peng Zou
contact affiliation	Peking University
contact email	zoupeng@pku.edu.cn
lab head
Lu Yan
contact affiliation	Peking University
contact email	yanlu18@pku.edu.cn
dataset submitter

iProX dataset URI