PXD022426-1

PXD022426 is an original dataset announced via ProteomeXchange.

Title	A tailored phosphoaspartate probe unravels CprR as a response regulator in Pseudomonas aeruginosa interkingdom signaling
Description	Pseudomonas aeruginosa is a difficult-to-treat Gram-negative bacterial pathogen causing life-threatening infections. Adaptive resistance (AR) to cationic peptide antibiotics such as polymyxin B impairs the therapeutic success. This self-protection is mediated by two component systems (TCS) consisting of a membrane-bound histidine kinase and an intracellular response regulator (RR). As phosphorylation of the key RR aspartate residue is transient during signaling and hydrolytically unstable, the study of these systems is challenging. Therefore, we applied a tailored reverse polarity chemical proteomic strategy to capture this transient modification and read-out RR phosphorylation in complex proteomes using a nucleophilic probe. An ideal trapping methodology was developed with a recombinant RR demonstrating the importance of fine-tuned acidic pH values to facilitate the attack on the aspartate carbonyl C-atom and prevent unproductive hydrolysis. Analysis of Bacillus subtilis and P. aeruginosa proteomes revealed the detection of multiple phosphoaspartate sites, which closely resembled the conserved RR sequence motif. With this validated strategy we dissected the signaling of dynorphin A, a human peptide stress hormone, which is sensed by P. aeruginosa to mediate AR. Intriguingly, our methodology identified CprR as an unprecedented RR in dynorphin A interkingdom signaling.
HostingRepository	PRIDE
AnnounceDate	2021-02-16
AnnouncementXML	Submission_2021-02-15_22:51:27.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Patrick Allihn
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562; scientific name: Bacillus subtilis; NCBI TaxID: 1423; scientific name: Pseudomonas aeruginosa PAO1; NCBI TaxID: 208964;
ModificationList	phosphorylated residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-09 03:24:21	ID requested
⏵ 1	2021-02-15 22:51:27	announced

Dataset with its publication pending

submitter keyword: Activity-based protein profiling, isotopically labeled tags, phosphorylation, response regulator, parallel reaction monitoring, adaptive resistance, antimicrobial peptide, B. subtilis, P. aeruginosa

Stephan Axel Sieber
contact affiliation	Technical University of Munich, Department of Chemistry, Chair of Organic Chemistry II, Lichtenbergstraße 4, 85748 Garching, Germany
contact email	stephan.sieber@tum.de
lab head
Patrick Allihn
contact affiliation	Technical University of Munich
contact email	patrick.allihn@tum.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022426
     1. Label: PRIDE project
     2. Name: A tailored phosphoaspartate probe unravels CprR as a response regulator in Pseudomonas aeruginosa interkingdom signaling