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ZIRC
ZFIN ID: ZDB-PUB-190118-3
Biochemical and structural analyses suggest that plasminogen activators coevolved with their cognate protein substrates and inhibitors
Jendroszek, A., Madsen, J.B., Chana-Muñoz, A., Dupont, D.M., Christensen, A., Panitz, F., Füchtbauer, E.M., Lovell, S.C., Jensen, J.K.
Date: 2019
Source: The Journal of biological chemistry   294(10): 3794-3805 (Journal)
Registered Authors:
Keywords: protein evolution, serine protease, serpin, substrate specificity, surface plasmon resonance (SPR)
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Evolution, Molecular*
  • Humans
  • Models, Molecular
  • Plasminogen Activator Inhibitor 1/metabolism*
  • Plasminogen Activators/antagonists & inhibitors
  • Plasminogen Activators/chemistry
  • Plasminogen Activators/metabolism*
  • Protein Binding
  • Protein Conformation
  • Urokinase-Type Plasminogen Activator/metabolism
PubMed: 30651349 Full text @ J. Biol. Chem.
ABSTRACT
Protein sequences of members of the plasminogen activation system are present throughout the entire vertebrate phylum. This important and well-described proteolytic cascade is governed by numerous protease-substrate and protease-inhibitor interactions whose conservation is crucial to maintaining unchanged protein function throughout evolution. The pressure to preserve protein-protein interactions may lead to either co-conservation or covariation of binding interfaces. Here, we combined covariation analysis and structure-based prediction to analyze the binding interfaces of urokinase (uPA):plasminogen activator inhibitor-1 (PAI-1) and uPA:plasminogen complexes. We detected correlated variation between the S3-pocket-lining residues of uPA and the P3 residue of both PAI-1 and plasminogen. These residues are known to form numerous polar interactions in the human uPA:PAI-1 Michaelis complex. To test the effect of mutations that correlate with each other and have occurred during mammalian diversification on protein-protein interactions, we produced uPA, PAI-1, and plasminogen from human and zebrafish to represent mammalian and nonmammalian orthologs. Using single amino acid point substitutions in these proteins, we found that the binding interfaces of uPA:plasminogen and uPA:PAI-1 may have coevolved to maintain tight interactions. Moreover, we conclude that although the interaction areas between protease-substrate and protease-inhibitor are shared, the two interactions are mechanistically different. Compared with a protease cleaving its natural substrate, the interaction between a protease and its inhibitor is more complex and involves a more fine-tuned mechanism. Understanding the effects of evolution on specific protein interactions may help further pharmacological interventions of the plasminogen activation system and other proteolytic systems.
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