Direct measurements of VEGF–VEGFR2 binding affinities reveal the coupling between ligand binding and receptor dimerization

King, Christopher R.; Hristova, Kalina

doi:10.1074/jbc.ra119.007737

Cited by 33 publications

(38 citation statements)

References 46 publications

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“…Altogether, this analysis provided further evidence that at least a subpopulation of VEGFR-2 on the surface of primary HDMECs exists in higher-order assembly states (dimers, oligomers, clusters or complexes) at rest, which most likely enhance VEGF binding to cells (16,17). This is however not exclusive, and some VEGF probably binds to VEGFR-2 monomers as well ( Fig.…”

Section: Co-imaging Of Vegfr-2 and Vegf Reveals That Vegf Binds To Anmentioning

confidence: 52%

“…Our study reveals the dynamic nanoscale organization of VEGFR-2 in its native environment on the surface of HDMECs, shedding light on the cell biophysical properties of this developmentally important receptor. In light of the higher avidity of VEGF to dimeric VEGFR-2 (16,17), this nanoscale organization most likely plays an important role in the responsiveness of ECs to VEGF. This nanoscale organization might also play a role in the responsiveness of ECs to shear stress (18)(19)(20).…”

Section: Discussionmentioning

confidence: 99%

“…The above experiments and analyses, especially the reduction in VEGFR-2 merging rate upon VEGF addition, suggest that a substantial fraction of VEGF binds to pre-existing VEGFR-2 dimers on the surface of HDMECs, with which it is expected to bind with higher avidity (16,17). To investigate this question further, we performed simultaneous 2-color TIRFM imaging of VEGFR-2 (labeled as above) and VEGF (conjugated with Atto488, which did not interfere with VEGF function (Fig.…”

Section: Co-imaging Of Vegfr-2 and Vegf Reveals That Vegf Binds To Anmentioning

confidence: 93%

“…However, as for some other RTKs (10)(11)(12)(13)(14), recent work provides evidence that VEGFR-2 can dimerize in the absence of ligand, and VEGF binding leads to dimer conformational changes that enable receptor trans-activation (15). Interestingly, VEGF has been observed to bind more avidly to VEGFR-2 pre-dimers (16,17). In addition, VEGFR-2 can be activated by fluid shear stress in a ligand-independent manner (18)(19)(20), raising the possibility of ligand-independent dimerization to enable activation, as dimerization is a prerequisite for the activation of many RTKs to relieve auto-inhibition (21).…”

Section: Introductionmentioning

confidence: 96%

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Heterogeneity in VEGF Receptor-2 mobility and organization on the endothelial cell surface leads to diverse activation models by VEGF

Rocha-Azevedo

Lee

Dasgupta

et al. 2019

Preprint

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Significance StatementVascular Endothelial Growth Factor Receptor 2 (VEGFR-2) is a critical receptor on the surface of endothelial cells (which line the blood vessels). It promotes the formation of new blood vessels in response to its ligand VEGF. Currently we know little about the organization and dynamics of this receptor on the cell surface, with multiple proposed models regarding its activation mechanism by VEGF. To fill this knowledge gap, we conducted a quantitative single-molecule imaging study of endogenous VEGFR-2 in primary endothelial cells, revealing the dynamic nanoscale organization of VEGFR-2 in its native environment. Our study also addresses the interplay between this organization, ligand binding and receptor activation, shedding light on the cell biophysical properties of this developmentally important receptor.

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Section: Co-imaging Of Vegfr-2 and Vegf Reveals That Vegf Binds To Anmentioning

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Section: Co-imaging Of Vegfr-2 and Vegf Reveals That Vegf Binds To Anmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 96%

See 2 more Smart Citations

Heterogeneity in VEGF Receptor-2 mobility and organization on the endothelial cell surface leads to diverse activation models by VEGF

Rocha-Azevedo

Lee

Dasgupta

et al. 2019

Preprint

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“…Switching between two dimerization modes of the transmembrane helix has recently been described as part of the activation mechanism of the EGF, VEGF and FGF receptors (26)(27)(28)(29). However, to date the role of TrkA-TMD dimerization in TrkA receptor activation has not been studied in detail.…”

Section: Structural Basis Of Trka Transmembrane Domain Dimerizationmentioning

confidence: 99%

Structural Basis of the Transmembrane Domain Dimerization in the Activation Mechanism of TrkA by NGF

Franco

Nadezhdin

Mineev

et al. 2019

Preprint

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Trk receptors are essential for the nervous system development. The molecular mechanism of TrkA activation by its ligand NGF is still unsolved. Recent data indicates that at endogenous levels most of TrkA is in an equilibrium monomer-dimer and the binding of NGF induces an increase of the dimer and oligomer forms of the receptor. An unsolved issue is the role of the transmembrane domain (TMD) in the dimerization of TrkA and the structural details of the TMD in the active dimer receptor. We found that TrkA-TMD can form dimers, identified the structural determinants of the dimer interface in the active receptor and validated this interface using site-directed mutagenesis together with functional and cell differentiation studies. Using in vivo crosslinking we identified a reordering of the extracellular juxtamembrane (JTM) region after ligand binding.Replacement of some residues in the JTM region with cysteine form ligand-independent active dimers and reveal a preferred dimer interface. In addition to that, insertion of leucine residues into the TMD helix induces a ligand-independent TrkA activation suggesting that a rotation of the TMD dimers could be behind TrkA activation by NGF. Altogether our data indicates that the transmembrane and juxtamembrane regions of the receptor play a key role in the dimerization and activation of TrkA by NGF.

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Methods | Ligand Binding to Receptor Tyrosine Kinases: Thermodynamic Cycles and Experimental Approaches

Light¹,

King²,

Hristova³

2021

Encyclopedia of Biological Chemistry III

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Direct measurements of VEGF–VEGFR2 binding affinities reveal the coupling between ligand binding and receptor dimerization

Cited by 33 publications

References 46 publications

Heterogeneity in VEGF Receptor-2 mobility and organization on the endothelial cell surface leads to diverse activation models by VEGF

Heterogeneity in VEGF Receptor-2 mobility and organization on the endothelial cell surface leads to diverse activation models by VEGF

Structural Basis of the Transmembrane Domain Dimerization in the Activation Mechanism of TrkA by NGF

Methods | Ligand Binding to Receptor Tyrosine Kinases: Thermodynamic Cycles and Experimental Approaches

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