Proteomics

Dataset Information

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A tailored phosphoaspartate probe unravels CprR as a response regulator in Pseudomonas aeruginosa interkingdom signaling


ABSTRACT: 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.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Escherichia Coli Pseudomonas Aeruginosa Pao1 Bacillus Subtilis

SUBMITTER: Patrick Allihn  

LAB HEAD: Stephan Axel Sieber

PROVIDER: PXD022426 | Pride | 2021-02-16

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
181103_SMH_181031_PA3.raw Raw
190421_PA_190411_1.raw Raw
190421_PA_190411_2.raw Raw
190421_PA_190411_3.raw Raw
190421_PA_190411_4.raw Raw
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