Improving the selectivity of engineered protease inhibitors: optimizing the P2 prime residue using a versatile cyclic peptide library

de Veer, Simon J., Wang, Conan, Harris, Jonathan M., Craik, David J. and Swedberg, Joakim E. (2015) Improving the selectivity of engineered protease inhibitors: optimizing the P2 prime residue using a versatile cyclic peptide library. Journal of Medicinal Chemistry, 58 20: 8257-8268. doi:10.1021/acs.jmedchem.5b01148


Author de Veer, Simon J.
Wang, Conan
Harris, Jonathan M.
Craik, David J.
Swedberg, Joakim E.
Title Improving the selectivity of engineered protease inhibitors: optimizing the P2 prime residue using a versatile cyclic peptide library
Journal name Journal of Medicinal Chemistry   Check publisher's open access policy
ISSN 0022-2623
1520-4804
Publication date 2015-09-22
Year available 2015
Sub-type Article (original research)
DOI 10.1021/acs.jmedchem.5b01148
Open Access Status Not yet assessed
Volume 58
Issue 20
Start page 8257
End page 8268
Total pages 12
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Subject 1313 Molecular Medicine
3002 Drug Discovery
Abstract Standard mechanism inhibitors are attractive design templates for engineering reversible serine protease inhibitors. When optimizing interactions between the inhibitor and target protease, many studies focus on the nonprimed segment of the inhibitor's binding loop (encompassing the contact beta-strand). However, there are currently few methods for screening residues on the primed segment. Here, we designed a synthetic inhibitor library (based on sunflower trypsin inhibitorI) for characterizing the P2' specificity of various serine proteases. Screening the library against 13 different proteases revealed unique P2' preferences for trypsin, ch-ymotrypsin, matriptase, plasmin, thrombin, four kallikrein-related peptidases, and several clotting factors. Using this information to modify existing engineered inhibitors yielded new variants that showed considerably improved selectivity, reaching up to 7000-fold selectivity over certain off-target proteases. Our study demonstrates the importance of the P2' residue in standard mechanism inhibition and unveils a new approach for screening P2' substitutions that will benefit future inhibitor engineering studies.
Formatted abstract
Standard mechanism inhibitors are attractive design templates for engineering reversible serine protease inhibitors. When optimizing interactions between the inhibitor and target protease, many studies focus on the nonprimed segment of the inhibitor’s binding loop (encompassing the contact β-strand). However, there are currently few methods for screening residues on the primed segment. Here, we designed a synthetic inhibitor library (based on sunflower trypsin inhibitor-1) for characterizing the P2′ specificity of various serine proteases. Screening the library against 13 different proteases revealed unique P2′ preferences for trypsin, chymotrypsin, matriptase, plasmin, thrombin, four kallikrein-related peptidases, and several clotting factors. Using this information to modify existing engineered inhibitors yielded new variants that showed considerably improved selectivity, reaching up to 7000-fold selectivity over certain off-target proteases. Our study demonstrates the importance of the P2′ residue in standard mechanism inhibition and unveils a new approach for screening P2′ substitutions that will benefit future inhibitor engineering studies.
Keyword Chemistry, Medicinal
Pharmacology & Pharmacy
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID 1059410
1026501
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 14 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 15 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Wed, 28 Oct 2015, 01:35:30 EST by Susan Allen on behalf of Office of the Vice-Chancellor