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Identification of N-terminal protein processing sites by chemical labeling mass spectrometry.

ABSTRACT: RATIONALE:Proteins undergo post-translational modifications and proteolytic processing that can affect their biological function. Processing often involves the loss of single residues. Cleavage of signal peptides from the N-terminus is commonly associated with translocation. Recent reports have suggested that other processing sites also exist. METHODS:The secreted proteins from S. aureus N315 were precipitated with trichloroacetic acid (TCA) and amidinated with S-methyl thioacetimidate (SMTA). Amidinated proteins were digested with trypsin and analyzed with a high-resolution orbitrap mass spectrometer. RESULTS:Sixteen examples of Staphylococcus aureus secretory proteins that lose an N-terminal signal peptide during their export were identified using this amidination approach. The N-termini of proteins with and without methionine were identified. Unanticipated protein cleavages due to sortase and an unknown protease were also uncovered. CONCLUSIONS:A simple N-terminal amidination based mass spectrometry approach is described that facilitates identification of the N-terminus of a mature protein and the discovery of unexpected processing sites.

SUBMITTER: Misal SA

PROVIDER: S-EPMC6522274 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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Identification of N-terminal protein processing sites by chemical labeling mass spectrometry.

Misal Santosh A SA Li Sujun S Tang Haixu H Radivojac Predrag P Reilly James P JP

Rapid communications in mass spectrometry : RCM 20190601 11

<h4>Rationale</h4>Proteins undergo post-translational modifications and proteolytic processing that can affect their biological function. Processing often involves the loss of single residues. Cleavage of signal peptides from the N-terminus is commonly associated with translocation. Recent reports have suggested that other processing sites also exist.<h4>Methods</h4>The secreted proteins from S. aureus N315 were precipitated with trichloroacetic acid (TCA) and amidinated with S-methyl thioacetim ...[more]

PMID: 30884002

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Dataset Information

Identification of N-terminal protein processing sites by chemical labeling mass spectrometry.

Publications

Identification of N-terminal protein processing sites by chemical labeling mass spectrometry.

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