bm0c00240_si_001.pdf (1.13 MB)
From In Silico to Experimental Validation: Tailoring Peptide Substrates for a Serine Protease
journal contribution
posted on 2020-04-01, 19:39 authored by Philip
Maximilian Knaff, Christian Kersten, Ramona Willbold, Carole Champanhac, Daniel Crespy, Rainer Wittig, Katharina Landfester, Volker MailänderSmart nanocarriers for the transport of drugs to tumor cells are
nowadays of great interest for treating cancer. The use of enzymatic
stimuli to cleave peptide-based drug nanocapsules for the selective
release of nanocapsule cargo in close proximity to tumor cells opens
new possibilities in cancer research. In the present work, we demonstrate
a methodology for finding and optimizing cleavable substrate sequences
by the type II transmembrane serine protease hepsin, which is highly
overexpressed in prostate cancer. The design and screening of combinatorial
libraries in silico against the binding cavity of hepsin allow the
identification of a panel of promising substrates with high-calculated
docking scores. In vitro screening verifies the predictions and showed
that all substrates are cleaved by hepsin with higher efficiency than
the literature known hepsin substrate RQLR↓VVGG. The introduction
of d-amino acids on a selected peptide with the highest catalytic
efficiency (kcat/Km) renders it resistant to cleavage by plasma or serum while
maintaining their susceptibility to hepsin.