PXD079543

PXD079543 is an original dataset announced via ProteomeXchange.

Title	ER-IP enables rapid isolation and proteomic characterization of the endoplasmic reticulum
Description	Isolation of the endoplasmic reticulum (ER) is essential for characterizing organelle-specific molecular processes, yet existing ER purification methods often require lengthy workflows and involve trade-offs between speed, specificity, and compatibility with downstream analyses. Here, we developed ER-IP, a rapid immunoaffinity-based method for ER isolation. ER-IP utilizes a Sec61β-3×HA-EGFP fusion protein to label ER membranes and enables affinity capture of ER fractions using anti-HA magnetic beads. We validated ER-IP through fluorescence microscopy, transmission electron microscopy, and biochemical analyses. The isolated fractions contained intact ER membrane structures and were enriched for canonical ER proteins. Compared with an ER-IP workflow incorporating a pre-clearing centrifugation step and a commercially available ER isolation kit, ER-IP substantially reduced processing time, enabling ER enrichment within approximately 15minutes while maintaining comparable recovery of ER-associated proteins. To further evaluate the molecular composition of ER-IP isolates, we performed quantitative proteomic analysis and identified 8,540 proteins. ER-IP fractions showed enrichment of proteins involved in protein translocation, protein folding, ER-associated degradation, lipid metabolism, and calcium homeostasis. In addition, the resulting proteomic dataset enabled identification of BAG2 as a previously unannotated ER-associated protein, illustrating the utility of ER-IP for discovery-oriented applications. Together, ER-IP provides a rapid and experimentally accessible approach for ER enrichment and downstream molecular characterization, expanding the toolkit available for investigating ER biology.
HostingRepository	iProX
AnnounceDate	2026-06-10
AnnouncementXML	Submission_2026-06-10_19:34:49.151.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wenmin Tian
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Ascend

Revision	Datetime	Status	ChangeLog Entry
0	2026-06-10 19:34:27	ID requested
⏵ 1	2026-06-10 19:34:49	announced

Dataset with its publication pending

submitter keyword: endoplasmic reticulum isolation, immunoaffinity purification,organelle enrichment

Yang Chen
contact affiliation	The Center for Precision Medicine Multi-omics Research (CPMMR)
contact email	chenyang1816185048@bjmu.edu.cn
lab head
Wenmin Tian
contact affiliation	The Center for Precision Medicine Multi-omics Research (CPMMR)
contact email	tianwenmin2019@126.com
dataset submitter

iProX dataset URI