An allosteric switch for pro-HGF/Met signaling using zymogen activator peptides

Kyle E Landgraf; Micah Steffek; Clifford Quan; Jeffrey Tom; Christine Yu; Lydia Santell; Henry R Maun; Charles Eigenbrot; Robert A Lazarus

doi:10.1038/nchembio.1533

An allosteric switch for pro-HGF/Met signaling using zymogen activator peptides

Nat Chem Biol. 2014 Jul;10(7):567-73. doi: 10.1038/nchembio.1533. Epub 2014 May 25.

Authors

Kyle E Landgraf¹, Micah Steffek², Clifford Quan³, Jeffrey Tom³, Christine Yu⁴, Lydia Santell³, Henry R Maun³, Charles Eigenbrot⁴, Robert A Lazarus³

Affiliations

¹ 1] Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA. [2].
² 1] Department of Structural Biology, Genentech, Inc., San Francisco, California, USA. [2].
³ Department of Early Discovery Biochemistry, Genentech, Inc., San Francisco, California, USA.
⁴ Department of Structural Biology, Genentech, Inc., San Francisco, California, USA.

PMID: 24859116
DOI: 10.1038/nchembio.1533

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.

MeSH terms

Allosteric Regulation / drug effects
Allosteric Site / drug effects
Amino Acid Sequence
Animals
CHO Cells
Catalytic Domain
Cell Movement / drug effects
Cell Survival / drug effects
Cricetulus
Gene Expression Regulation
Hepatocyte Growth Factor / chemistry
Hepatocyte Growth Factor / genetics
Hepatocyte Growth Factor / metabolism*
Humans
Models, Molecular
Molecular Mimicry
Molecular Sequence Data
Peptide Library
Peptides / chemical synthesis
Peptides / pharmacology*
Protein Binding
Protein C / chemistry
Protein C / genetics
Protein C / metabolism
Protein Engineering
Protein Precursors / chemistry
Protein Precursors / genetics
Protein Precursors / metabolism*
Protein Structure, Tertiary
Proto-Oncogene Proteins / chemistry
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism
Proto-Oncogene Proteins c-met / chemistry
Proto-Oncogene Proteins c-met / genetics
Proto-Oncogene Proteins c-met / metabolism*
Signal Transduction / drug effects*

Substances

Peptide Library
Peptides
Protein C
Protein Precursors
Proto-Oncogene Proteins
macrophage stimulating protein
pro-hepatocyte growth factor
Hepatocyte Growth Factor
MET protein, human
Proto-Oncogene Proteins c-met