Biophysical Studies of the Induced Dimerization of Human VEGF Receptor 1 Binding Domain by Divalent Metals Competing with VEGF-A

PLoS One. 2016 Dec 12;11(12):e0167755. doi: 10.1371/journal.pone.0167755. eCollection 2016.

Abstract

Angiogenesis is tightly regulated through the binding of vascular endothelial growth factors (VEGFs) to their receptors (VEGFRs). In this context, we showed that human VEGFR1 domain 2 crystallizes in the presence of Zn2+, Co2+ or Cu2+ as a dimer that forms via metal-ion interactions and interlocked hydrophobic surfaces. SAXS, NMR and size exclusion chromatography analyses confirm the formation of this dimer in solution in the presence of Co2+, Cd2+ or Cu2+. Since the metal-induced dimerization masks the VEGFs binding surface, we investigated the ability of metal ions to displace the VEGF-A binding to hVEGFR1: using a competition assay, we evidenced that the metals displaced the VEGF-A binding to hVEGFR1 extracellular domain binding at micromolar level.

MeSH terms

  • Binding Sites
  • Cations, Divalent / pharmacology*
  • Humans
  • Molecular Docking Simulation*
  • Protein Binding / drug effects
  • Protein Multimerization*
  • Vascular Endothelial Growth Factor A / chemistry*
  • Vascular Endothelial Growth Factor A / metabolism
  • Vascular Endothelial Growth Factor Receptor-1 / chemistry*
  • Vascular Endothelial Growth Factor Receptor-1 / metabolism

Substances

  • Cations, Divalent
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factor Receptor-1

Grants and funding

This work was supported by: 1- Centre National de la Recherche Scientifique (CNRS), http://www.cnrs.fr: JFG MRS NGE SBr PC BS MBL BG WQL FH NI SBo MV IB. The CNRS had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. 2- Universite Paris Descartes (UPD), http://www.parisdescartes.fr: JFG MRS NGE SBr PC BS MBL BG WQL FH SBo MV IB. The UPD had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. 3- Fondation ARC pour la recherche sur le cancer (ARC), http://www.recherche-cancer.net/. The ARC had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. 4- Agence Nationale pour la Recherche (ANR), http://www.agence-nationale-recherche.fr/projet-anr/?solr=run&tx_lwmsuivibilan_pi2%5BCODE%5D=ANR-10-BLAN-1533. The ANR had validated and funded the research project, but it had no role in data collection and analysis, decision to publish, or preparation of the manuscript.