{"database":"panorama","file_versions":[],"scores":null,"additional":{"omics_type":["Proteomics"],"submitter":["Laura Eck"],"species":["Staphylococcus Aureus"],"full_dataset_link":["https://panoramaweb.org/ClpP-Trap-PRM.url"],"submitter_email":["laura.eck@tum.de"],"submitter_affiliation":["TUM"],"sample_protocol":[""],"repository":["PanoramaPublic"],"data_protocol":[""],"description_synonyms":["ClpP, Proteolytic Enzyme, D17Wsu160e, RP11-508D10.1, 2-Amino-3-hydroxypropionic acid, Esteroproteases, Tnfsf5, Hydrolase, Techniques, Regions, Method, Proteases, Analysis, acting on D-amino acid peptides, Staphlococcus pyogenes citreus, Catalytic Core, T-BAM, Mass Spectrum Analysis, type 5 acid phosphatase, Eubacteria, tartrate-resistant acid ATPase, Analyses, pre-mortem, Subunit, Staphilococcus aureus, endopeptidase Ti activity, Homeostases, Serine Proteinases, Subunits, Micrococcus aureus, procedures, Proteolytic, Proteinases, peptide hydrolase activity, Bacteria Woese et al. 2024, TRAP3, T-cell antigen Gp39, Tudor repeat associator with PCTAIRE-2, Bacteria <bacteria>, Methodological Studies, 2-amino-3-hydroxypropanoic acid, Prokaryotae, Trapping, Procaryotae, TNFSF5, T5ap, MGC - 45012, Catalytic Subunits, CD40L, Proteinase, Tnfrsf5, Arts, Cd40l, familial, Procedure, Spectrum Analysis, hCD40L, Spectroscopy, 3-Hydroxyalanine, Catalytic Regions, peptidase activity, Domains, IMD3, prokaryotes, TNF-related activation protein, Region, Ly-62, Domain, FPF, Industrial, Serine, Industrial Arts, Fishing, NKTL, Spectrometry, Reactive Site, AW743063, Methodological, AI326936, ATP-dependent Clp protease, Methodological Study, living, Staphylococcus pyogenes aureus, TrATPase, IGM, CD40LG, L Serine, Reactive Sites, L-Serine, Prokaryota, Proteolytic Enzymes, AU019820, Procedures, TRAP, p50, Peptidomics, Ly62, TAPK, Protein Homeostasis, Gene, Protease, eubacteria, Spectrum Analyses, Active Site, TR-AP, autosomal dominant, Genetic Interventions., DFNB81, GP39, Sites, Studies, Gene Products, Mass, Bacteriobiota, Peptide Hydrolase, CD40-L, Technique, Reactive, Genetic Intervention, Mass Spectroscopy, gp39, CD154, Site, Genetic, CATC4, GKLP, Serine Proteinase, protease Ti activity, Monera, Proteostases, Bacteria (ex Cavalier-Smith 1987), Clp protease, Study, PCTAIRE2-binding protein, Enzyme, MGC:45012, Catalytic, Catalytic Subunit, TEIF, Trap, caseinolytic protease activity, acting on L-amino acid peptides, Cores, HIGM1, Serine Protein Hydrolases, fungi, acting on peptide bonds, Staphylococus aureus, Catalytic Cores, hydrolase, C78062, Catalytic Region, Alkyne, P105, NTKL, protease activity, bacteria, PCTAIRE2BP, Catalytic Site, Protein Homeostases, Proteins, Active Sites, Bp50, ACP5, SPENCDI, Peptide, Cell, Core, Serin, MS, Protein, periodic FEVER, Peptidases, Active, techniques, Catalytic Sites, Acetylenes, Mass Spectrum Analyses, Acetylenic Compounds, Intervention, Mass Spectrum, Exhibit, Staphylococcus aureus subsp. anaerobius, Homeostasis, Catalytic Domains, Protein Hydrolases, Engineering, Streptococcus aureus, Protein Gene Products, PRLTS3, Gene Proteins, Peptidase, Micrococcus pyogenes, proteinase activity, prokaryote, Serine Protease, Interventions, TRACP, methodology"],"name_synonyms":["CD40L, AU019820, P105, NTKL, ClpP, TRAP, p50, PCTAIRE2BP, Ly62, Tnfrsf5, TAPK, Cd40l, Bp50, D17Wsu160e, ACP5, SPENCDI, TR-AP, RP11-508D10.1, hCD40L, Tnfsf5, DFNB81, GP39, IMD3, TNF-related activation protein, CD40-L, Ly-62, gp39, T-BAM, CD154, Action, type 5 acid phosphatase, tartrate-resistant acid ATPase, CATC4, GKLP, protease Ti activity, endopeptidase Ti activity, NKTL, AW743063, AI326936, ATP-dependent Clp protease, PRLTS3, Clp protease, PCTAIRE2-binding protein, TRAP3, T-cell antigen Gp39, Generic Action., Tudor repeat associator with PCTAIRE-2, MGC:45012, TEIF, Trap, caseinolytic protease activity, TrATPase, IGM, HIGM1, CD40LG, TNFSF5, TRACP, T5ap, MGC - 45012, C78062"],"additional_accession":[]},"is_claimable":false,"name":"It’s a trap: ClpP substrates caught in action","description":"Protein 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.","dates":{"publication":"Thu Oct 03 00:00:00 GMT+01:00 2024"},"accession":"PXD047469","cross_references":{"TAXONOMY":["1280"]}}