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PXD047274-2

PXD047274 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleA Photocrosslinking Probe to Capture the Substrates of Caseinolytic Protease P
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 remain 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. After enrichment using an alkyne handle, substrate deconvolution by mass spectrometry (MS) is performed. 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 for the discovery of serine protease substrates beyond genetic engineering.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_07:00:43.597.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterNina Bach
SpeciesList scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationListacetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentQ Exactive HF; Orbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-11-27 06:55:20ID requested
12024-10-04 04:50:35announced
22024-10-22 07:00:44announced2024-10-22: Updated project metadata.
Publication List
10.1002/anie.202409220;
Keyword List
submitter keyword: S. aureus, substrates
Contact List
Stephan Sieber
contact affiliationTUM School of Natural Sciences, Department Biosciences, Chair of Organic Chemistry II, Center for Functional Protein Assemblies (CPA), Technical University Munich (TUM), Ernst-Otto-Fischer Str. 8, Garching, 85748, Germany
contact emailstephan.sieber@tum.de
lab head
Nina Bach
contact affiliationTU München
contact emailnina.bach@tum.de
dataset submitter
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Dataset FTP location
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