Hydrophobic tagging-assisted N-termini enrichment (HYTANE) for in-depth N-terminome analysis

The analysis of protein N-termini is of great importance for understanding the protein function and elucidating the proteolytic processing. Herein, we develop a negative enrichment strategy, termed as hydrophobic tagging-assisted N-termini enrichment (HYTANE) to achieve a global N-terminome analysis. The HYTANE strategy showed a high efficiency in hydrophobic tagging and C18 material-assisted depletion using bovine serum albumin (BSA) as the sample. Furthermore, this strategy was also applied to N-termini profiling from S. cerevisiaecell lysates and enabled the identification of 1096 protein N-termini, representing the largest N-terminome dataset of S. cerevisiae. The identified N-terminal peptides accounted for 99% of all identified peptides and no deficiency in acidic and histidine-containing N-terminal peptides was observed. The presented HYTANE strategy is therefore a highly selective, efficient and unbiased strategy for the large scale N-terminome analysis.