Engineering high-efficiency matriptase substrates using E. coli display for applications in prodrug activation.

ABSTRACT: Proteases play a crucial role in biological functions such as tumor progression and tissue homeostasis. Recently, protease-activated prodrugs have gained attention for their potential to enhance selectivity in tumor-targeted therapies. In this study, we report the engineering of substrate sequences for matriptase, a protease overexpressed in tumors and previously explored for prodrug activation in vivo. A peptide library containing millions of potential substrates was displayed on Escherichia coli, and flow cytometric sorting was used to isolate improved substrates based on cleavage efficiency. Hits were ranked by flow cytometry, and the top substrates exhibited k_cat/K_M values over 40-fold higher than previously reported sequences. These substrates were further evalua

SUBMITTER: Mestre Borras A 

PROVIDER: S-EPMC12272249 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature