Exploiting Selective Position Labeling to Unveil the Hidden Complexity of Terminomics

Selective protein terminal labeling has become essential for system-wide studies of proteolytic mechanisms in disease. These methods enable precise tracking of cleavage dynamics, protease interactions, and cellular networks, offering transformative potential for proteolytic event analysis. This review explores recent advances in N-/C-terminal modification strategies, specifically for the applications in terminomics—the field focused on protein termini characterization. While protein termini provide valuable insights into functional proteome states, their low abundance in complex samples demands highly selective labeling approaches. We evaluate modern chemical and chemoenzymatic methods that leverage engineered chemical reactivity thresholds or enzymatic precision for site-specific modifications. Emerging strategies show enhanced substrate adaptability, reaction efficiency, and workflow compatibility, enabling broader applications in terminome studies.