an N-terminal acetylated peptide enrichment method using CNBr-activated sepharose resin
Protein N-terminal acetylation (N(α)-acetylation) is observed widely from prokaryotes to eukaryotes. It gains increased importance in biological field, due to its multiple roles in many aspects of the protein life, such as assembly, stability, activity, and location. Today, mass spectrometry (MS) has demonstrated its unprecedented ability in a variety of proteome-wide studies including the N(α)-acetylome. The N(α)-acetylated peptides are usually immerged into the massive amount of regular peptides and are further suppressed due to their reduced positive charge states during analysis; therefore, an efficient exploration of the N(α)-acetylome necessitates a specific enrichment method. Here we describe a protocol for N(α)-acetylated peptide enrichment using CNBr-activated sepharose resin, which has proved to be simple, sensitive, and highly reproducible.
Methods in Molecular Biology, 2013, Vol 981, p. 47-56
Acetylation; Chromatography, Liquid; Cyanogen Bromide; HeLa Cells; Humans; Peptides; Proteome; Reproducibility of Results; Sensitivity and Specificity; Sepharose; Tandem Mass Spectrometry