title: An allosteric switch for pro-HGF/Met signaling using zymogen activator peptides.
abstract: Stimulation of hepatocyte growth factor (HGF) signaling through the Met receptor is an attractive approach for promoting tissue repair and preventing fibrosis. Using structure-guided peptide phage display combined with an activity-based sorting strategy, we engineered allosteric activators of zymogen-like pro-HGF to bypass proteolytic activation and reversibly stimulate pro-HGF signaling through Met. Biochemical, structural and biological data showed that zymogen activator peptides (ZAPtides) potently and selectively bind the activation pocket within the serine protease-like β-chain of pro-HGF and display titratable activation of pro-HGF-dependent Met signaling, leading to cell survival and migration. To further demonstrate the versatility of our ZAPtide platform, we identified allosteric activators for pro-macrophage stimulating protein and a zymogen serine protease, Protein C, which also provides evidence for target selectivity. These studies reveal that ZAPtides use molecular mimicry of the trypsin-like N-terminal insertion mechanism and establish a new paradigm for selective pharmacological activation of plasminogen-related growth factors and zymogen serine proteases.
authors.affiliation: 1] Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA. [2].
authors.forename: Kyle E
authors.initials: KE
authors.lastname: Landgraf
authors.affiliation: 1] Department of Structural Biology, Genentech, Inc., San Francisco, California, USA. [2].
authors.forename: Micah
authors.initials: M
authors.lastname: Steffek
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
authors.forename: Clifford
authors.initials: C
authors.lastname: Quan
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Tom
authors.affiliation: Department of Structural Biology, Genentech, Inc., San Francisco, California, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Yu
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
authors.forename: Lydia
authors.initials: L
authors.lastname: Santell
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
authors.forename: Henry R
authors.initials: HR
authors.lastname: Maun
authors.affiliation: Department of Structural Biology, Genentech, Inc., San Francisco, California, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Eigenbrot
authors.affiliation: Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Lazarus
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D000495:Allosteric Site / Q000187:drug effects; D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D020134:Catalytic Domain; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003412:Cricetulus; D005786:Gene Expression Regulation; D017228:Hepatocyte Growth Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011485:Protein Binding; D011486:Protein C / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015202:Protein Engineering; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D019151:Peptide Library; D010455:Peptides; D011486:Protein C; D011498:Protein Precursors; D011518:Proto-Oncogene Proteins; C082332:macrophage stimulating protein; C086013:pro-hepatocyte growth factor; D017228:Hepatocyte Growth Factor; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met