PXD024470

PXD024470 is an original dataset announced via ProteomeXchange.

Title	Succinylome-controlled antibiotic resistance in MRSA - Succinylome
Description	A 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.
HostingRepository	MassIVE
AnnounceDate	2021-06-14
AnnouncementXML	Submission_2021-06-14_15:27:55.799.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Matthew Monroe
SpeciesList	scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationList	Carbamidomethyl; TMT6plex; Oxidation
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2021-03-02 11:38:29	ID requested
⏵ 1	2021-06-14 15:27:56	announced

no publication

submitter keyword: proteomics, antibiotic resistance, MRSA, SucC, succinyl-lysine, succinylome

James P. O'Gara
contact affiliation	National University of Ireland, Galway
contact email	jamesp.ogara@nuigalway.ie
lab head
Geremy Clair
contact affiliation	Pacific Northwest National Laboratory
contact email	geremy.clair@pnnl.gov
lab head
Matthew Monroe
contact affiliation	Pacific Northwest National Laboratory
contact email	matthew.monroe@pnnl.gov
dataset submitter

MassIVE dataset URI

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