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PXD047469

PXD047469 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleIt’s a trap: ClpP substrates caught in action
DescriptionProtein homeostasis in bacteria is regulated by proteases such as the tetradecameric caseinolytic protease P (ClpP). Although substrates of ClpP have been successfully deciphered in genetically engineered cells, methods which directly trap processed proteins within native cells are elusive. Here we introduce an in situ trapping strategy which utilizes trifunctional probes that bind to the active site serine of ClpP and capture adjacent substrates with an attached photocrosslinking moiety. Substrate deconvolution by mass spectrometry (MS) is accomplished after enrichment using an alkyne handle. We show that our two traps bind substoichiometrically to ClpP, retain protease activity, exhibit unprecedented selectivity for Staphylococcus aureus ClpP in living cells and capture numerous known and novel substrates. The exemplary validation of trapped hits using a targeted proteomics approach confirmed the fidelity of this technology. In conclusion, we provide a novel chemical platform suited to discover serine protease substrates beyond genetic engineering.
HostingRepositoryPanoramaPublic
AnnounceDate2024-10-03
AnnouncementXMLSubmission_2024-10-03_10:56:44.662.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterLaura Eck
SpeciesList scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationListNo PTMs are included in the dataset
InstrumentOrbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-12-03 00:11:35ID requested
12024-10-03 10:56:45announced
Publication List
Angew. Chem. Int. Ed. 2024, e202409220
Keyword List
submitter keyword: S. aureus, ClpP, protease substrate trapping, ClpP substrates, parallel reaction monitoring
Contact List
Stephan Sieber
contact affiliationTUM
contact emailstephan.sieber@tum.de
lab head
Laura Eck
contact affiliationTUM
contact emaillaura.eck@tum.de
dataset submitter
Full Dataset Link List
Panorama Public dataset URI