Projects per year
Abstract
We developed a simple and rapid method to enrich protein N-terminal peptides, in which the protease TrypN is first employed to generate protein N-terminal peptides without Lys or Arg and internal peptides with two positive charge sat their N termini, and then, the N-terminal peptides with or without N-acetylation are separated from the internal peptides by strong cation exchange chromatography according to a retention model based on the charge/orientation of peptides. This approach was applied to20 μg of human HEK293T cell lysate proteins to profile theN-terminal proteome. On average, 1550 acetylated and200 unmodified protein N-terminal peptides were successfully identified in a single LC/MS/MS run with less than 3% contamination with internal peptides, even when we accepted only canonical protein N termini registered in the Swiss-Prot database. Because this method involves only two steps, protein digestion and chromatographic separation, without the need for tedious chemical reactions, it should be useful for comprehensive profiling of protein N termini, including proteo forms with neo-Ntermini.
Original language | English |
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Article number | 100003 |
Number of pages | 9 |
Journal | Molecular & Cellular Proteomics (MCP) |
Volume | 20 |
Early online date | 23 Nov 2020 |
DOIs | |
Publication status | Published - 1 Jan 2021 |
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Dive into the research topics of 'Isolation of acetylated and unmodified protein N-Terminal peptides by strong cation exchange chromatographic separation of TrypN-digested peptides'. Together they form a unique fingerprint.Projects
- 3 Finished
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`Core Funding for the Wellcome Trust Centre for Cell Biology¿, Research Enrichment, Public Engagement
Tollervey, D. (Principal Investigator)
1/12/18 → 1/06/22
Project: Research
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Wellcome Centre for Cell Biology
Tollervey, D. (Principal Investigator)
1/12/16 → 1/12/21
Project: Research
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Protein structures in the context of time and space by mass spectrometry
Rappsilber, J. (Principal Investigator)
1/06/14 → 31/05/21
Project: Research