Addition of a c-myc epitope tag within the VEGF protein does not affect in vitro biological activity

Biochem Cell Biol. 2001;79(1):107-12.

Abstract

The overexpression of vascular endothelial growth factor (VEGF) has been strongly implicated in diseases involving neovascularization. VEGF exists in as many as six different isoforms, each showing a unique pattern of tissue distribution and activity. To investigate the effect of individual VEGF isoform overexpression in neovascular disease models, we inserted c-myc epitope tags into the three VEGF isoforms expressed in retinal pigment epithelial cells, VEGF121, VEGF165, and VEGF189. We found that the 12-amino acid insertion between the receptor binding and heparin binding domains did not affect VEGF transcription, translation, or secretion. In addition, VEGF isoforms containing the c-myc epitope tag were able to stimulate endothelial cell proliferation as efficiently as non-tagged VEGF isoforms and they could be individually identified by Western blotting and immunocytochemistry using the c-myc epitope specific monoclonal antibody 9E10.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cell Line
  • DNA Primers
  • Endothelial Growth Factors / chemistry*
  • Endothelial Growth Factors / genetics
  • Endothelial Growth Factors / metabolism*
  • Epitopes / chemistry*
  • Humans
  • Immunohistochemistry
  • Lymphokines / chemistry*
  • Lymphokines / genetics
  • Lymphokines / metabolism*
  • Proto-Oncogene Proteins c-myc / chemistry*
  • RNA, Messenger / genetics
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors

Substances

  • DNA Primers
  • Endothelial Growth Factors
  • Epitopes
  • Lymphokines
  • Proto-Oncogene Proteins c-myc
  • RNA, Messenger
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Vascular Endothelial Growth Factors