PXD065905

PXD065905 is an original dataset announced via ProteomeXchange.

Title	A high-efficiency Azide-A-DSBSO cross-linking-MS protocol for identifying thousands of in vivo protein interactions in single day
Description	Enrichable and mass spectrometry (MS)-cleavable crosslinkers have gained popularity in crosslinking-MS (XL-MS) in the past decade. The azide-tagged, acid-cleavable disuccinimidyl bis-sulfoxide (DSBSO) crosslinker offers both these advantages, enabling deep XL-MS interactomics studies in intact cells. However, current DSBSO XL-MS workflows take several days, which limits the experimental throughput. Here, we developed an in-cell DSBSO XL-MS protocol that allows sample ready for MS analysis within 8 hours. By systematically optimizing digestion, click chemistry enrichment, LC parameters, and MS fragmentation, we increased the number of crosslink spectrum matches (CSMs) from single-shot experiments more than twofold compared to previous single-shot measurements. This allowed us to efficiently generate deep protein-protein interaction datasets from intact human mitochondria, Bacillus subtilis, and HEK293T cells.
HostingRepository	PRIDE
AnnounceDate	2025-11-21
AnnouncementXML	Submission_2025-11-21_08:58:05.968.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Pin-Lian Jiang
SpeciesList	scientific name: Bacillus subtilis; NCBI TaxID: NEWT:1423; scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-07-08 06:44:06	ID requested
⏵ 1	2025-11-21 08:58:06	announced

10.1002/ANIE.202518355;

submitter keyword: high-thoughput,cross-linking mass spectrometry, DSBSO

Fan Liu
contact affiliation	Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP)
contact email	liu.fan(at)fmp-berlin.de
lab head
Pin-Lian Jiang
contact affiliation	Leibniz-Forschungsinstitut für Molekulare Pharmakologie
contact email	jiang@fmp-berlin.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD065905
     1. Label: PRIDE project
     2. Name: A high-efficiency Azide-A-DSBSO cross-linking-MS protocol for identifying thousands of in vivo protein interactions in single day