Dataset Information

P.aeruginosa PAO1+Pht Tandem Mass Tag (TMT)-based quantitative proteomic analysis

ABSTRACT: To study the changes in the proteomics profile of P. aeruginosa PAO1 upon Pht addition, TMT was used as the labeling strategy for comparative quantitative proteomic analysis.

ORGANISM(S): Pseudomonas Aeruginosa

SUBMITTER: Liang Yang

PROVIDER: PXD032384 | iProX | Fri Mar 18 00:00:00 GMT 2022

REPOSITORIES: iProX

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Publications

The AhR ligand phthiocol and vitamin K analogs as Pseudomonas aeruginosa quorum sensing inhibitors.

Jia Tianyuan T Liu Dongjing D Bi Xianbiao X Li Menglu M Cai Zhao Z Fu Jiapeng J Liu Zhi Z Wu Pengyao P Ke Xue X Jia Aiqun A Zhang Guoliang G Li Guobao G Yang Liang L

Frontiers in microbiology 20220914

The aryl hydrocarbon receptor (AhR) protein senses microbial-secreted metabolites to trigger the host's innate immune system. The Pseudomonas quinolone signal (PQS) and Mycobacterium tuberculosis (MTb) metabolite phthiocol (Pht) are both ligands of AhR with similar chemical structures. As PQS is an essential quorum-sensing molecule that regulates a wide range of virulence factors in Pseudomonas aeruginosa, we hypothesized that Pht and its analogs are potential P. aerugino ...[more]

PMID: 36187967

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Project description:Fractionation is essential to achieving deep proteome coverage for sample multiplexing experiments where currently up to 18 samples can be analyzed concurrently. However, peptide fractionation (i.e., upstream of LC-MS/MS analysis) with a liquid chromatography system constrains sample processing as only a single sample can be fractionated at once. Here, we highlight the use of spin column-based methods which permit multiple multiplexed samples to be fractionated simultaneously. These methods require only a centrifuge and eliminate the need for a dedicated liquid chromatography system. We investigate peptide fractionation with strong anion exchange (SAX) and high-pH reversed phase (HPRP) spin columns, as well as a combination of both. In two separate experiments, we acquired deep proteome coverage (>8000 quantified proteins), while starting with <25 μg of protein per channel. Our datasets showcase the proteome alterations in two human cell lines resulting from treatment with inhibitors acting on the ubiquitin-proteasome system. We recommend this spin column-based peptide fractionation strategy for high-throughput screening applications or whenever a liquid chromatograph is not readily available. SIGNIFICANCE: Fractionation is a means to achieve deep proteome coverage for global proteomics analysis. Typical liquid chromatography systems may be a prohibitive expense for many laboratories. Here, we investigate prefractionation with strong anion exchange (SAX) and high-pH reversed phase (HPRP) spin columns, as well as a combination of both, as peptide fractionation methods. These spin columns have advantages over liquid chromatography systems, which include relative affordability, higher throughput capability, no carry over, and fewer potential instrument-related malfunctions. In two separate experiments, we acquired deep proteome coverage (>8000 quantified proteins), thereby showing the utility of each or a combination of both spin columns for global proteome analysis.

Dataset Information

P.aeruginosa PAO1+Pht Tandem Mass Tag (TMT)-based quantitative proteomic analysis

Publications

The AhR ligand phthiocol and vitamin K analogs as <i>Pseudomonas aeruginosa</i> quorum sensing inhibitors.

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