Quantitative profiling of protease specificity.

摘要: Proteases are an important class of enzymes, whose activity is central to many physiologic and pathologic processes. Detailed knowledge protease specificity key understanding their function. Although methods 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"; set amino acid sequences that uniquely represent 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, which accounts for number substrates relative Using these advances greatly facilitates elucidation selectivity between family. The study provides insight into degree cleft defines recognition, thus providing basis overcoming major challenges field proteolysis: 1) development highly selective probes studying proteases overlapping specificities, 2) distinguishing targeted proteolysis from bystander proteolytic events.