Efficient activation of the lymphangiogenic growth factor VEGF-C requires the C-terminal domain of VEGF-C and the N-terminal domain of CCBE1

Sci Rep. 2017 Jul 7;7(1):4916. doi: 10.1038/s41598-017-04982-1.


The collagen- and calcium-binding EGF domains 1 (CCBE1) protein is necessary for lymphangiogenesis. Its C-terminal collagen-like domain was shown to be required for the activation of the major lymphangiogenic growth factor VEGF-C (Vascular Endothelial Growth Factor-C) along with the ADAMTS3 (A Disintegrin And Metalloproteinase with Thrombospondin Motifs-3) protease. However, it remained unclear how the N-terminal domain of CCBE1 contributed to lymphangiogenic signaling. Here, we show that efficient activation of VEGF-C requires its C-terminal domain both in vitro and in a transgenic mouse model. The N-terminal EGF-like domain of CCBE1 increased VEGFR-3 signaling by colocalizing pro-VEGF-C with its activating protease to the lymphatic endothelial cell surface. When the ADAMTS3 amounts were limited, proteolytic activation of pro-VEGF-C was supported by the N-terminal domain of CCBE1, but not by its C-terminal domain. A single amino acid substitution in ADAMTS3, identified from a lymphedema patient, was associated with abnormal CCBE1 localization. These results show that CCBE1 promotes VEGFR-3 signaling and lymphangiogenesis by different mechanisms, which are mediated independently by the two domains of CCBE1: by enhancing the cleavage activity of ADAMTS3 and by facilitating the colocalization of VEGF-C and ADAMTS3. These new insights should be valuable in developing new strategies to therapeutically target VEGF-C/VEGFR-3-induced lymphangiogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • COS Cells
  • Calcium-Binding Proteins / genetics*
  • Calcium-Binding Proteins / metabolism
  • Chlorocebus aethiops
  • Endothelial Cells / metabolism*
  • Endothelial Cells / pathology
  • Gene Expression Regulation
  • HEK293 Cells
  • Humans
  • Lymphangiogenesis / genetics*
  • Lymphatic Vessels / metabolism*
  • Lymphatic Vessels / pathology
  • Lymphedema / genetics*
  • Lymphedema / metabolism
  • Lymphedema / pathology
  • Mice
  • Mice, Transgenic
  • NIH 3T3 Cells
  • Protein Domains
  • Protein Precursors / genetics
  • Protein Precursors / metabolism
  • Proteolysis
  • Signal Transduction
  • Tumor Suppressor Proteins / genetics*
  • Tumor Suppressor Proteins / metabolism
  • Vascular Endothelial Growth Factor C / genetics*
  • Vascular Endothelial Growth Factor C / metabolism
  • Vascular Endothelial Growth Factor Receptor-3 / genetics
  • Vascular Endothelial Growth Factor Receptor-3 / metabolism


  • CCBE1 protein, human
  • Calcium-Binding Proteins
  • Protein Precursors
  • Tumor Suppressor Proteins
  • VEGFC protein, human
  • Vascular Endothelial Growth Factor C
  • Vascular Endothelial Growth Factor Receptor-3