Dataset Information

Extensive arginine phosphorylation in Staphylococcus aureus is influences by Stp1

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Fusion Lumos, Orbitrap Fusion, Orbitrap Exploris 480

ORGANISM(S): Escherichia Coli Staphylococcus Aureus Bacillus Subtilis Subsp. Subtilis Str. 168

SUBMITTER: Nadine Prust

LAB HEAD: Simone Lemeer

PROVIDER: PXD026981 | Pride | 2022-06-09

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20180428_OR14_Ag1_Prust001_SA_2mg_Bsubtilis_IMAC_1.raw	Raw
	20180428_OR14_Ag1_Prust001_SA_2mg_Bsubtilis_IMAC_2.raw	Raw
	20180428_OR14_Ag1_Prust001_SA_2mg_Bsubtilis_IMAC_3.raw	Raw
	20190215_OR15_Ag1_Prust001_SA_2mg_SdrE_1_IMAC_exp7.raw	Raw
	20190215_OR15_Ag1_Prust001_SA_2mg_SdrE_2_IMAC_exp7.raw	Raw

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Publications

Widespread Arginine Phosphorylation in Staphylococcus aureus.

Prust Nadine N van Breugel Pieter C PC Lemeer Simone S

Molecular & cellular proteomics : MCP 20220412 5

Arginine phosphorylation was only recently discovered to play a significant and relevant role in the Gram-positive bacterium Bacillus subtilis. In addition, arginine phosphorylation was also detected in Staphylococcus aureus, suggesting a widespread role in bacteria. However, the large-scale analysis of protein phosphorylation, and especially those that involve a phosphoramidate bond, comes along with several challenges. The substoichiometric nature of protein phosphorylation requires proper enr ...[more]

PMID: 35421590

Publication: 1/2

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