title: 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.
abstract: METHODS
  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 antiangiogenic activity on human umbilical vein cells. 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.
  
  
  RESULTS
  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 dependent signaling was the target of TCMP bioactivity. Angiogenesis was inhibited in vivo in zebrafish embryos treated with 500 nM TCMP.
  
  
  CONCLUSIONS
  Our results demonstrate that cystine-knot miniproteins present in mature tomato fruits are endowed with antiangiogenic 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.
authors.affiliation: Department of Biotechnology, University of Verona, Verona, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Treggiari
authors.affiliation: Department of Biotechnology, University of Verona, Verona, Italy.
authors.forename: Gianni
authors.initials: G
authors.lastname: Zoccatelli
authors.affiliation: Department of Biotechnology, University of Verona, Verona, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Molesini
authors.affiliation: Department of Medicine, Section of Internal Medicine, University of Verona, Verona, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Degan
authors.affiliation: CRA-ORL Council for Agriculture Research and Economics, Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi, Italy.
authors.forename: Giuseppe Leonardo
authors.initials: GL
authors.lastname: Rotino
authors.affiliation: CRA-ORL Council for Agriculture Research and Economics, Research Unit for Vegetable Crops, Montanaso Lombardo, Lodi, Italy.
authors.forename: Tea
authors.initials: T
authors.lastname: Sala
authors.affiliation: L.U.R.M. University Laboratory for Medical Research, University of Verona, Verona, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Cavallini
authors.affiliation: Department of Medicine, Cardiovascular Medicine Division, Brigham and Women's Hospital, Boston, USA.
authors.forename: Calum A
authors.initials: CA
authors.lastname: MacRae
authors.affiliation: Department of Medicine, Section of Internal Medicine, University of Verona, Verona, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Minuz
authors.affiliation: Department of Biotechnology, University of Verona, Verona, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Pandolfini
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D057771:Cystine-Knot Miniproteins / Q000302:isolation & purification / Q000494:pharmacology*; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D005638:Fruit / Q000737:chemistry; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018551:Solanum lycopersicum / Q000737:chemistry*; D009569:Nitric Oxide / Q000096:biosynthesis*; D010766:Phosphorylation; D010940:Plant Proteins / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor / Q000502:physiology*; D015027:Zebrafish
chemical_list: D020533:Angiogenesis Inhibitors; D057771:Cystine-Knot Miniproteins; D010940:Plant Proteins; D009569:Nitric Oxide; D040262:Receptors, Vascular Endothelial Growth Factor; D051057:Proto-Oncogene Proteins c-akt
keywords: Angiogenesis; Cell migration; Cystine-knot protein; Tomato fruit; VEGFA signaling