Structural Determinants of Vascular Endothelial Growth factor-D Receptor Binding and Specificity

Blood. 2011 Feb 3;117(5):1507-15. doi: 10.1182/blood-2010-08-301549. Epub 2010 Dec 8.

Abstract

Vascular endothelial growth factors (VEGFs) and their tyrosine kinase receptors (VEGFR-1-3) are central mediators of angiogenesis and lymphangiogenesis. VEGFR-3 ligands VEGF-C and VEGF-D are produced as precursor proteins with long N- and C-terminal propeptides and show enhanced VEGFR-2 and VEGFR-3 binding on proteolytic removal of the propeptides. Two different proteolytic cleavage sites have been reported in the VEGF-D N-terminus. We report here the crystal structure of the human VEGF-D Cys117Ala mutant at 2.9 Å resolution. Comparison of the VEGF-D and VEGF-C structures shows similar extended N-terminal helices, conserved overall folds, and VEGFR-2 interacting residues. Consistent with this, the affinity and the thermodynamic parameters for VEGFR-2 binding are very similar. In comparison with VEGF-C structures, however, the VEGF-D N-terminal helix was extended by 2 more turns because of a better resolution. Both receptor binding and functional assays of N-terminally truncated VEGF-D polypeptides indicated that the residues between the reported proteolytic cleavage sites are important for VEGF-D binding and activation of VEGFR-3, but not of VEGFR-2. Thus, we define here a VEGFR-2-specific form of VEGF-D that is angiogenic but not lymphangiogenic. These results provide important new insights into VEGF-D structure and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Proliferation
  • Cells, Cultured
  • Crystallography, X-Ray
  • Humans
  • Hydrogen Bonding
  • Immunoenzyme Techniques
  • Immunoprecipitation
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / metabolism*
  • Mutagenesis, Site-Directed
  • Mutation / genetics
  • Protein Binding
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Vascular Endothelial Growth Factor C / chemistry
  • Vascular Endothelial Growth Factor C / genetics
  • Vascular Endothelial Growth Factor C / metabolism
  • Vascular Endothelial Growth Factor D / chemistry*
  • Vascular Endothelial Growth Factor D / genetics
  • Vascular Endothelial Growth Factor D / metabolism*
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism*
  • Vascular Endothelial Growth Factor Receptor-3 / metabolism*

Substances

  • Recombinant Proteins
  • VEGFC protein, human
  • Vascular Endothelial Growth Factor C
  • Vascular Endothelial Growth Factor D
  • Vascular Endothelial Growth Factor Receptor-2
  • Vascular Endothelial Growth Factor Receptor-3

Associated data

  • PDB/2XV7