NAA10 serves as the catalytic subunit for both the NatA and N-terminal acetyltransferase E (NatE) complexes. To investigate the impact of the NAA10p.S37P mutation on N-terminal acetylation in COs, we employed the HUNTER strategy[小刘7.1][MS7.2] to enrich acetylated N-terminal peptides. Briefly, free amines of internal (non-N-terminal) tryptic peptides were modified with undecanal after trypsin digestion. This increased non-N-terminal peptide hydrophobicity, allowing them to be retained on a reverse-phase chromatography column, while N-terminal acetylation peptides were not retained. These peptides were further purified by C18 solid-phase extraction. After drying in a SpeedVac concentrator, the samples were reconstituted in 0.1% TFA and subjected to LC–MS/MS analysis.