Development of a novel peptide inhibitor of subtilisin BPN′. Issue 11 (22nd September 2022)

• Record Type: Journal Article
• Authors: Ishida, Kohki; Shimizu, Makoto; Wakasugi, Ayumi; Matsui, Yuko; Nakamura, Akira; Kojima, Shuichi
• Other Names: Rizo Josep guestEditor.
• Is Part Of: FEBS open bio. Volume 12:Issue 11(2022)
• Page Start: 2057
• Page End: 2064
• Publication Date: 2022-09-22
• Subjects: disulfide bond -- inhibitory peptide -- phage display -- protease inhibitor -- protein–peptide interactions -- subtilisin BPN′; Molecular biology -- Periodicals; Cytology -- Periodicals; Life sciences -- Periodicals; Biological Science Disciplines -- Periodicals; Molecular Biology -- Periodicals; Cell Biology -- Periodicals; Cytology; Life sciences; Molecular biology; Periodicals; 572.805
• DOI: 10.1002/2211-5463.13481
• Languages: English
• ISSNs: 2211-5463

Abstract

Abstract : Proteinaceous protease inhibitors can strongly and specifically inhibit cognate proteases, but their use as pharmaceuticals is limited by their size. As such, the development of effective protease peptide inhibitors would be beneficial for biochemical studies and drug discovery. In this study, we applied a phage display system to select subtilisin BPN′‐binding peptides and evaluated their inhibitory activities against subtilisin BPN′. A 12mer peptide with an intramolecular disulfide bond inhibited subtilisin BPN′ ( K i value of 13.0 nm ). Further mutational analyses of the peptide resulted in the development of a short peptide inhibitor against subtilisin BPN′ that showed high inhibitory activity and binding affinity ( K i value of 0.30 nm ). This activity was found to be derived from the conformational rigidity caused by the intramolecular disulfide bond and the small residue at the P1′ site and from the interaction of the P4 and P6′ residues with subtilisin BPN′. Abstract : We identified several potential inhibitory peptides for subtilisin BPN′, a typical serine protease, by phage display method. Of these, a 12mer peptide, named ASIP, showed inhibitory activity with an inhibition constant ( K i ) of 13 nm against subtilisin BPN′. Subsequent mutation analyses on this peptide led to the development of an improved inhibitory peptide with K i of 0.3 nm .