title: Efficient degradation-aided selection of protease inhibitors by phage display.
abstract: In this report, we describe a novel phage display strategy for the identification of dedicated protease inhibiting peptides, based on degradation-aided enrichment of protease resistant phages. Phages were directly incubated with a range of phage-degrading proteases, after which non-degraded phages were used for the next selection round. For proteinase-K we identified after only four selection rounds a peptide (VLIMPVLLGIPLLC) that inhibits proteinase-K activity with an inhibition constant of 4 microM. In analogy, we identified a peptide capable of inhibiting substrate degradation by cathepsin-S (VWNCERITISRLIN), which showed functional inhibition of cathepsin-S induced sprouting of endothelial cells. We envision that the pursued strategy of degradation-aided selection of protease inhibitors (DASPI) represents an effective approach in the design of new protease inhibitors but also of new strategies to render gene and drug vectors protease resistant.
authors.affiliation: Leiden/Amsterdam Centre for Drug Research, Division of Biopharmaceutics, Leiden, The Netherlands. L.J.A.C.Hawinkels@LUMC.nl
authors.forename: Lukas J A C
authors.initials: LJ
authors.lastname: Hawinkels
authors.forename: Sabine M W
authors.initials: SM
authors.lastname: van Rossenberg
authors.forename: Eveline S M
authors.initials: ES
authors.lastname: de Jonge-Muller
authors.forename: Tom J M
authors.initials: TJ
authors.lastname: Molenaar
authors.forename: Chantal C M
authors.initials: CC
authors.lastname: Appeldoorn
authors.forename: Theo J C
authors.initials: TJ
authors.lastname: van Berkel
authors.forename: Cornelis F M
authors.initials: CF
authors.lastname: Sier
authors.forename: Erik A L
authors.initials: EA
authors.lastname: Biessen
mesh_terms: D015195:Drug Design; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D011480:Protease Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D019151:Peptide Library; D010455:Peptides; D011480:Protease Inhibitors