Tracking N-terminal protein processing from the Golgi to the chromatophore of a rhizarian amoeba.
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ABSTRACT:
SUBMITTER: Meinnel T
PROVIDER: S-EPMC9237673 | biostudies-literature | 2022 Jun
REPOSITORIES: biostudies-literature
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Tracking N-terminal protein processing from the Golgi to the chromatophore of a rhizarian amoeba.
Plant physiology 20220601 3
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Project description: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.
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