PXD026981

PXD026981 is an original dataset announced via ProteomeXchange.

Title	Extensive arginine phosphorylation in Staphylococcus aureus is influences by Stp1
Description	Ever since the identification of the first protein-arginine kinase, McsB, in B. subtilis arginine phosphorylation gained more attention. However, the analysis of phosphorylations especially phosphormaidates comes along with several challenges. The sub-stoichiometric nature of protein phosphorylation requires proper enrichment strategies prior to LC-MS/MS analysis and the acid instability of phosphoramidates was long though to impede those enrichments. Further good spectral quality is required which can be reduced by the presence of neutral losses of phosphoric acid upon higher energy collision induced dissociation. Adaptation of common enrichment strategies to less acidic conditions and the use of electron-transfer dissociation allowed the identification of more than 200 pArg sites in B. subtilis and S. aureus so far. Here we show that pArg is stable enough for commonly used Fe3+-IMAC enrichment followed by LC-MS/MS and that HCD is still the gold standard for phosphoproteomics. By profiling a serine/ threonine kinase (Stk1) and phosphatase (Stp1) mutant from a methicillin-resistant S. aureus mutant library, we identified 1062 pArg sites and thus the most comprehensive arginine phosphoproteome to date. Using synthetic phosphoarginine peptides we validated the presence and localization of arginine phosphorylation in S. aureus and lastly, we could show that Stp1 is influencing the arginine phosphoproteom without being a protein-arginine phosphatase.
HostingRepository	PRIDE
AnnounceDate	2022-06-09
AnnouncementXML	Submission_2022-06-09_07:07:44.684.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nadine Prust
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Bacillus subtilis subsp. subtilis str. 168; NCBI TaxID: 224308; scientific name: Staphylococcus aureus; NCBI TaxID: 1280;
ModificationList	phosphorylated residue
Instrument	Orbitrap Fusion Lumos; Orbitrap Exploris 480; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-06-28 21:24:28	ID requested
⏵ 1	2022-06-09 07:07:45	announced

Prust N, van der Laarse S, van den Toorn HWP, van Sorge NM, Lemeer S, In-Depth Characterization of the Staphylococcus aureus Phosphoproteome Reveals New Targets of Stk1. Mol Cell Proteomics, 20():100034(2021) [pubmed]

Prust N, van Breugel PC, Lemeer S, Widespread Arginine Phosphorylation in Staphylococcus aureus. Mol Cell Proteomics, 21(5):100232(2022) [pubmed]

submitter keyword: arginine phosphorylation, Staphylococcus aureus, Stp1, phosphorylation, fragmentation

Simone Lemeer
contact affiliation	Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands
contact email	S.M.Lemeer@uu.nl
lab head
Nadine Prust
contact affiliation	Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands
contact email	n.prust@uu.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD026981
     1. Label: PRIDE project
     2. Name: Extensive arginine phosphorylation in Staphylococcus aureus is influences by Stp1