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PXD063970-1

PXD063970 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleRapid photo-crosslinking in living cells reveals protein-nucleic acid dynamics on a timescale of minutes
DescriptionThe activation of chemical reactions in living cells using ultraviolet (UV) light enables the interrogation of biomolecules in their native environment with photoreactive probes or crosslinking reagents. Although numerous photo-crosslinking approaches have been successfully employed, they often suffer from common limitations, including low reaction yields, the need for long exposure times, and irradiation-induced cellular damage from heat, desiccation, or side reactions. We recently showed that 365 nm light-emitting diodes (LEDs) enable rapid, biorthogonal protein-DNA crosslinking in living cells, incurring minimal photodamage. Here we generalize this approach and demonstrate that high-intensity, longwave UV light reduces the irradiation time for in-cell photo-crosslinking reactions by up to 1000-fold, allowing protein-drug, protein-protein, protein-DNA and protein-RNA interactions to be fixed within seconds. Benchmarking this rapid photo-activation for the analysis of RNA-interacting proteomes responding to RNA-binding drugs or UV-induced RNA damage, we demonstrate both qualitative and quantitative advantages of controlled, high-intensity UV irradiation, uncovering emergent experimental opportunities that were previously inaccessible to light-activated chemistry in intact cells and tissues.
HostingRepositoryMassIVE
AnnounceDate2026-04-23
AnnouncementXMLSubmission_2026-04-23_04:17:49.298.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterBernhard Kuster
SpeciesList scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentOrbitrap Eclipse; timsTOF HT; Orbitrap Astral
Dataset History
RevisionDatetimeStatusChangeLog Entry
02025-05-15 03:57:13ID requested
12026-04-23 04:17:49announced
Publication List
no publication
Keyword List
submitter keyword: photo-crosslinking, high-intensity UV irradiation, UVEN, in-cell photo-activation, photo-affinity ligation (PAL), protein-protein crosslinking, SDA, photo-activatable nucleotides, protein-DNA crosslinking, 4-thiothymidine, protein-RNA crosslinking, 4-thiouridine, crosslinking kinetics, DatasetType:Proteomics
Contact List
Bernhard Kuster
contact affiliationChair of Proteomics and Bioanalytics, Technical University of Munich
contact emailkuster@tum.de
lab head
Bernhard Kuster
contact affiliationChair of Proteomics and Bioanalytics, Technical University of Munich
contact emailproteomics.kusterlab@gmail.com
dataset submitter
Full Dataset Link List
MassIVE dataset URI
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://massive-ftp.ucsd.edu/v09/MSV000097903/