HUNTER: Sensitive Automated Characterization of Proteolytic Systems by N Termini Enrichment from Microscale Specimen

Proteolysis occurs at low frequency in the cellular environment. Protein N termini reveal essential mechanisms associated with cellular functions, and are useful indicators to track dysfunctional regulation of proteins and pathways in diseases. N terminomics has so far relied on labor-intensive methods, which require relatively large starting sample amounts rendering it ill-suited for high-throughput systems biology studies. Here, we describe protocols for the first scalable and automatable method for sensitive enrichment and identification of N termini from minute samples.