Artificial Zymogen\nBased on Protein–Polymer\nHybrids

This study explores the synthesis and application of\nartificial\nzymogens using protein–polymer hybrids to mimic the controlled\nenzyme activation observed in natural zymogens. Pro-trypsin (pro-TR)\nand pro-chymotrypsin (pro-CT) hybrids were engineered by modifying\nthe surfaces of trypsin (TR) and chymotrypsin (CT) with cleavable\npeptide inhibitors utilizing surface-initiated atom transfer radical\npolymerization. These hybrids exhibited 70 and 90% reductions in catalytic\nefficiency for pro-TR and pro-CT, respectively, due to the inhibitory\neffect of the grafted peptide inhibitors. The activation of pro-TR\nby CT and pro-CT by TR resulted in 1.5- and 2.5-fold increases in\nenzymatic activity, respectively. Furthermore, the activated hybrids\ntriggered an enzyme activation cascade, enabling amplification of\nactivity through a dual pro-protease hybrid system. This study highlights\nthe potential of artificial zymogens for therapeutic interventions\nand biodetection platforms by harnessing enzyme activation cascades\nfor precise control of catalytic activity.