Mapping Cell Surface Proteolysis with Plasma Membrane-Targeted Subtiligase.
Mapping Cell Surface Proteolysis with Plasma Membrane-Targeted Subtiligase.
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DOI：
10.1007/978-1-0716-2124-0_6
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发表时间：
2022
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N terminomics methods combine selective isolation of protein N-terminal peptides with mass spectrometry (MS)-based proteomics for global profiling of proteolytic cleavage sites. However, traditional N terminomics workflows require cell lysis before N-terminal enrichment and provide poor coverage of N termini derived from cell surface proteins. Here, we describe application of subtiligase-TM, a plasma membrane-targeted peptide ligase, for selective biotinylation of cell surface N termini, enabling their enrichment and analysis by liquid chromatography-tandem MS (LC-MS/MS). This method provides increased coverage of and specificity for cell surface N termini and is compatible with existing quantitative LC-MS/MS workflows.
