Quantitative profiling of protease specificity.

摘要: Abstract Proteases comprise an important class of enzymes, whose activity is central to many physiologic and pathologic processes. Detailed knowledge protease specificity key understanding their function. Although methodologies have been developed profile specificities proteases, few the diversity quantitative grasp necessary fully define a protease, both in terms substrate numbers catalytic efficiencies. We concept “selectome”, which defines set substrates that uniquely represents protease. applied it two closely related members Matrixin family – MMP-2 MMP-9 by using phage display coupled with Next Generation Sequencing information theory-based data analysis. also derived measure specificity, accounts for relative efficiencies substrates. Using these advances greatly facilitates uncovering selectivity between families provides insight into degree contribution cleft protein recognition, thus providing basis overcoming major challenges field proteolysis: 1) development highly selective probes inhibitors studying proteases overlapping specificities, 2) distinguishing targeted proteolysis from bystander proteolytic events.