title: Measuring net protease activities in biological samples using selective peptidic inhibitors.
abstract: The measurement of activities from individual proteases in biological samples is difficult because of the numerous proteases, their overlapping activities, and the lack of specific substrates. We applied selective protease inhibitors based on bicyclic peptides (>2000-fold selective over related proteases) to block individual proteases, allowing the quantification of their net activities. In protease mixtures, activity contributions of the serine proteases plasma kallikrein and urokinase-type plasminogen activator (uPA) were accurately quantified. In a tumor extract, we could quantify uPA activity. Because bicyclic peptide inhibitors toward virtually any protease can be generated by phage display, the approach should be applicable to any protease.
authors.affiliation: Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Lisa
authors.initials: L
authors.lastname: Pollaro
authors.forename: Philippe
authors.initials: P
authors.lastname: Diderich
authors.forename: Alessandro
authors.initials: A
authors.lastname: Angelini
authors.forename: Silvia
authors.initials: S
authors.lastname: Bellotto
authors.forename: Hermann
authors.initials: H
authors.lastname: Wegner
authors.forename: Christian
authors.initials: C
authors.lastname: Heinis
mesh_terms: D006801:Humans; D007610:Kallikreins / Q000097:blood; D007700:Kinetics; D009369:Neoplasms / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000032:analysis / Q000097:blood; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011480:Protease Inhibitors / Q000737:chemistry*; D014568:Urokinase-Type Plasminogen Activator / Q000097:blood
chemical_list: D019151:Peptide Library; D010455:Peptides; D011480:Protease Inhibitors; D010447:Peptide Hydrolases; D007610:Kallikreins; D014568:Urokinase-Type Plasminogen Activator