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A CpxA Phosphatase Inhibitor Activates CpxRA and is an Effective Treatment for Uropathogenic Escherichia coli in a Murine Model of Infection

ABSTRACT: For label-free quantitative proteomics, cell pellets from 5 independent experiments were subjected for protein extraction followed by LC-MS/MS. LC-MS/MS analysis was performed on a Dionex UltiMate 3000 (Thermo Fisher Scientific) system connected to a Q-Exactive HF-X mass spectrometer (Thermo Fisher Scientific). Peptide spectrum matching of MS/MS spectra was searched against the UniProt E. coli CFT073-UPEC database (TaxID:199310) using the Sequest algorithm within Proteome Discoverer v 2.4 software (Thermo Fisher Scientific). The Sequest database search was performed with the following parameters: trypsin enzyme cleavage specificity, 2 possible missed cleavages, 10 ppm mass tolerance for precursor ions, 0.02 Da mass tolerance for fragment ions. Search parameters permitted dynamic modification of methionine oxidation (+15.9949 Da) and static modification of carbamidomethylation (+57.0215 Da) on cysteine. Peptide assignments from the database search were filtered to a 1% False Discovery Rate. The relative label-free quantitative and comparative among the samples were performed using the Minora algorithm and Proteome Discoverer 2.4 software. Differential abundance of proteins between conditions was defined as a 1.5-fold change in relative abundance with an adjusted p-value < 0.05.

INSTRUMENT(S): Dionex UltiMate 3000 connected to Q Exactive HF-X mass spectrometer

ORGANISM(S): Escherichia Coli Cft073 Taxid:199310

SUBMITTER: Stanley M Spinola

PROVIDER: MSV000088484 | MassIVE | Wed Dec 01 08:13:00 GMT 2021

REPOSITORIES: MassIVE

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