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PXD024470

PXD024470 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleSuccinylome-controlled antibiotic resistance in MRSA - Succinylome
DescriptionA sucC and a sucD mutant from the Nebraska Transposon Mutant Library (NTML, Strain NE569 and NE1770) were found to be susceptible to cefoxitin and oxacillin when other mutations in TCA cycle genes did not affect their resistance to these antibiotics. The SucC strain (Strain NE569) accumulated Succinyl CoA, which is known to result in higher succinylation rate. We compared the proteome of the sucC mutant in exponential and stationary stages to the one of the WT strain JE2. The global proteome of the following strains were analyzed by TMT-labelled proteomics: Early exponential JE2 (3Hrs), Exponential JE2 (4Hrs), Exponential SucC (4Hrs), Stationary JE2 (10Hrs), Stationary SucC (12Hrs). For each condition, the bacterial pellet was collected and washed twice with PBS. The proteins were extracted by bead beating, 1000 ug of proteins were reduced with DTT, alkylated with IAA and digested with 1:50 (trypsin:protein, w:w) for 3 Hr at 37 C. 100ug of peptides were labelled with TMT11Plex reagents. The samples were pooled and enriched for Succinyl-lysine containing peptides using the PTMScan Succinyl-Lysine Motif Enrichment Kit following the supplier's instructions. The retained fraction was cleaned-up by C18 solid phase extraction. The peptides were concentrated in 15 ul and 5 ul were analyzed by reverse-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS) using a Waters nanoEquity UPLC system coupled with a QExactive HF-X mass spectrometer.
HostingRepositoryMassIVE
AnnounceDate2021-06-14
AnnouncementXMLSubmission_2021-06-14_15:27:55.799.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterMatthew Monroe
SpeciesList scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationListCarbamidomethyl; TMT6plex; Oxidation
InstrumentQ Exactive HF-X
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-03-02 11:38:29ID requested
12021-06-14 15:27:56announced
Publication List
no publication
Keyword List
submitter keyword: proteomics, antibiotic resistance, MRSA, SucC, succinyl-lysine, succinylome
Contact List
James P. O'Gara
contact affiliationNational University of Ireland, Galway
contact emailjamesp.ogara@nuigalway.ie
lab head
Geremy Clair
contact affiliationPacific Northwest National Laboratory
contact emailgeremy.clair@pnnl.gov
lab head
Matthew Monroe
contact affiliationPacific Northwest National Laboratory
contact emailmatthew.monroe@pnnl.gov
dataset submitter
Full Dataset Link List
MassIVE dataset URI
Dataset FTP location
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