A cystine-knot miniprotein from tomato fruit inhibits endothelial cell migration and angiogenesis by affecting vascular endothelial growth factor receptor (VEGFR) activation and nitric oxide production

Treggiari, D., Zoccatelli, G., Molesini, B., Degan, M., Rotino, G.L., Sala, T., Cavallini, C., MacRae, C.A., Minuz, P., Pandolfini, T.
Molecular Nutrition & Food Research   59(11): 2255-66 (Journal)
Registered Authors
MacRae, Calum A.
Angiogenesis, Cell migration, Cystine-knot protein, Tomato fruit, VEGFA signaling
MeSH Terms
  • Angiogenesis Inhibitors/pharmacology*
  • Animals
  • Cell Movement/drug effects*
  • Cells, Cultured
  • Cystine-Knot Miniproteins/isolation & purification
  • Cystine-Knot Miniproteins/pharmacology*
  • Endothelial Cells/drug effects*
  • Endothelial Cells/physiology
  • Fruit/chemistry
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Nitric Oxide/biosynthesis*
  • Phosphorylation
  • Plant Proteins/pharmacology*
  • Proto-Oncogene Proteins c-akt/metabolism
  • Receptors, Vascular Endothelial Growth Factor/physiology*
  • Solanum lycopersicum/chemistry*
  • Zebrafish
26255647 Full text @ Mol. Nutr. Food Res.
Cystine-knot miniproteins are bioactive molecules with a broad range of potential therapeutic applications. Recently, it was demonstrated that two tomato cystine-knot miniproteins (TCMPs) exhibit in vitro anti-angiogenic activity on human umbilical vein cells (HUVEC). The aim of the present study was to investigate the effects of a fruit-specific cystine-knot miniprotein of tomato on in vitro endothelial cell migration and in vivo angiogenesis using a zebrafish model.
The cystine-knot protein purified from tomato fruits using gel filtration LC and RP- HPLC, inhibited cell migration when tested at 200 nM using the wound-healing assay, and reduced nitric oxide formation probed by 4-amino-5-methylamino-27-difluorofluoscescin diacetate. RT-PCR and western blot analyses demonstrated that vascular endothelium growth factor A (VEGFA)-dependent signaling was the target of TCMP bioactivity. Angiogenesis was inhibited in vivo in zebrafish embryos treated with 500 nM TCMP.
Our results demonstrate that cystine-knot miniproteins present in tomato mature fruits are endowed with anti-angiogenic activity in vitro and in vivo. These molecules may confer beneficial effects to tomato dietary intake, along with lycopene and other antioxidants. Further investigation is warranted to explore the potential of these compounds as model scaffolds for the development of new drugs. This article is protected by copyright. All rights reserved.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes