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

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.

Ionophore Ability of Carnosine and Its Trehalose Conjugate Assists Copper Signal in Triggering Brain-Derived Neurotrophic Factor and Vascular Endothelial Growth Factor Activation In Vitro

“…Namely, both extracellular domains show similar structural features with the presence of histidine residues which contribute to the cognate ligand binding and can also act as chelating anchors for d-block metal ions such as copper(II) and zinc(II). Consistently, a recent report shows that human VEGFR1 domain 2 crystallizes in the presence of Cu 2+ as a dimer that is formed via metal ion interactions and interlocked hydrophobic surfaces, recalling the dimerization effect induced by VEGF interaction with its receptor [135]. A competition test puts in evidence that Cu 2+ is able to displace VEGF from its VEGFR1 extracellular domain at micromolar level.…”

“…Treatment of cells for 24 h resulted in an increase in VEGF level in cells of up to 147 ± 7%. Altogether, the findings highlight the role of copper in connecting BDNF and VEGF release and expression; this interplay is further supported by the ability of Cu 2+ to interact with the VEGFR1 domain2, inducing dimer formation [135] and recalling the effect of VEGF interaction with its receptor. Furthermore, it is noteworthy that both the specific receptors of BDNF and VEGF belong to the same family of RTKs and their ectodomains share a significant copper(II) ion affinity with subsequent activation of extracellular metal signaling.…”

Disrupting VEGF–VEGFR1 Interaction: De Novo Designed Linear Helical Peptides to Mimic the VEGF13-25 Fragment

“…To avoid false positive results due to possible contamination with metals or peptide aggregation [ 26 , 27 ], the screening was repeated at the same dose of 30 μM, but in the presence of EDTA, for those compounds having inhibition values above 60% ( Table 2 ). The new experiments performed in the presence of EDTA led to slightly lower inhibition percentage values, especially for peptides with high Trp content.…”

Vascular Endothelial Growth Factor Receptor, fms-Like Tyrosine Kinase-1 (Flt-1), as a Novel Binding Partner for SARS-CoV-2 Spike Receptor-Binding Domain

“…The 3D structure of spike-RBD and sFlt-1 proteins were recovered from the UniProt 3D structure PBD database (PDB DOI: 10.2210/pdb6W41/PDB And:10.2210/pdb4CKV/PDB). sFlt-1 and spike-RBD 3D structures were compared by referring to the published structure of the proteins (17,18). Additionally, sequence alignment of amino acid residues of sFlt-1 with either spike RBD or ACE2 and neuropilin-1 which are known binding partners of spike RBD was performed using Clustal Omega multiple sequence alignment tools.…”

“…Ligand was prepared by retrieving d2-sFlt1 3D-structure from protein data bank (PDB)-4CKV deposited in Research Collaboratory for Structural Bioinformatics (RCSB) (18). Detection and selection of the reaction roots optimized, then choosing and setting the number of active torsions using ligand preparation tools of Auto Dock 4.…”