Projects per year
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.
`Core Funding for the Wellcome Trust Centre for Cell Biology¿, Research Enrichment, Public Engagement
1/12/18 → 1/06/22
1/06/14 → 31/05/21