title: Design of Specific Serine Protease Inhibitors Based on a Versatile Peptide Scaffold: Conversion of a Urokinase Inhibitor to a Plasma Kallikrein Inhibitor.
abstract: All serine proteases hydrolyze peptide bonds by the same basic mechanism and have very similar active sites, in spite of the fact that individual proteases have different physiological functions. We here report a strategy for designing high-affinity and high-specificity serine protease inhibitors using a versatile peptide scaffold, a 10-mer peptide, mupain-1 (CPAYSRYLDC). Mupain-1 was previously reported as a specific inhibitor of murine urokinase-type plasminogen activator (Ki = 0.55 μM) without measurable affinity to plasma kallikrein (Ki > 1000 μM). On the basis of a structure-based rational design, we substituted five residues of mupain-1 and converted it to a potent plasma kallikrein inhibitor (Ki = 0.014 μM). X-ray crystal structure analysis showed that the new peptide was able to adapt a new set of enzyme surface interactions by a slightly changed backbone conformation. Thus, with an appropriate re-engineering, mupain-1 can be redesigned to specific inhibitors of other serine proteases.
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Department of Molecular Biology and Genetics, Aarhus University , Aarhus, 8000, Denmark.
authors.forename: Peng
authors.initials: P
authors.lastname: Xu
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences , Fuzhou, 350002, China.
authors.forename: Mingming
authors.initials: M
authors.lastname: Xu
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences , Fuzhou, 350002, China.
authors.forename: Longguang
authors.initials: L
authors.lastname: Jiang
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Department of Molecular Biology and Genetics, Aarhus University , Aarhus, 8000, Denmark.
authors.forename: Qinglan
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne National Laboratory, Argonne, Illinois 60439, United States.
authors.forename: Zhipu
authors.initials: Z
authors.lastname: Luo
authors.affiliation: Synchrotron Radiation Research Section, Macromolecular Crystallography Laboratory, National Cancer Institute, Argonne National Laboratory, Argonne, Illinois 60439, United States.
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Dauter
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences , Fuzhou, 350002, China.
authors.forename: Mingdong
authors.initials: M
authors.lastname: Huang
authors.affiliation: Danish-Chinese Centre for Proteases and Cancer, Department of Molecular Biology and Genetics, Aarhus University , Aarhus, 8000, Denmark.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Andreasen
mesh_terms: D001798:Blood Proteins / Q000138:chemical synthesis* / Q000494:pharmacology*; D015195:Drug Design; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000737:chemistry*; D010456:Peptides, Cyclic / Q000737:chemistry / Q000494:pharmacology; D020842:Plasma Kallikrein / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000737:chemistry; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis* / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D001798:Blood Proteins; D010455:Peptides; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; C530075:mupain-1; C031220:urokinase inhibitor; D020842:Plasma Kallikrein