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Parallel reaction monitoring analysis of target proteins

ABSTRACT: Parallel reaction monitoring (PRM) analysis was used to confirm changes in abundance of the target proteins (AceF, SodA, YopD, YopE, GroEL, TerE, and OsmY2),. Equal amounts of proteins from the culture supernatants and cell pellets were separated by SDS-PAGE, and bands corresponding to each of the target proteins were subjected to in-gel digestion. The peptide samples for the target proteins were then analyzed by mass spectrometry, and three peptides from each target protein were selected for the PRM analysis. The selection criterion for the peptides was a sequence length of 6–15 amino acids, while peptides with any modifications or missed cleavage sites were excluded. The peak area of three b-/y-ions from each peptide was used to calculate the peptide ratio, and the ratios of three peptides were used to determine the target protein ratio in different samples.

ORGANISM(S): Yersinia Pestis Co92

SUBMITTER: Ruifu Yang

PROVIDER: PXD020061 | iProX | Wed Dec 16 00:00:00 GMT 2020

REPOSITORIES: iProX

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Secretome and Comparative Proteomics of Yersinia pestis Identify Two Novel E3 Ubiquitin Ligases That Contribute to Plague Virulence.

Cao Shiyang S Chen Yuling Y Yan Yanfeng Y Zhu Songbiao S Tan Yafang Y Wang Tong T Song Yajun Y Deng Haiteng H Yang Ruifu R Du Zongmin Z

Molecular & cellular proteomics : MCP 20210222

Plague is a zoonotic disease that primarily infects rodents via fleabite. Transmission from flea to host niches requires rapid adaption of Yersinia pestis to the outer environments to establish infection. Here, quantitative proteome and secretome analyses of Y. pestis grown under conditions mimicking the two typical niches, i.e., the mammalian host (Mh) and the flea vector (Fv), were performed to understand the adaption strategies of this deadly pathogen. A secretome of Y. pestis containing 308 ...[more]

PMID: 33631294

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