Proteomic identification of VEGF-dependent protein enrichment to membrane caveolar-raft microdomains in endothelial progenitor cells

Anastasia Chillà, Francesca Magherini, Francesca Margheri, Anna Laurenzana, Tania Gamberi, Luca Bini, Laura Bianchi, Giovanna Danza, Benedetta Mazzanti, Simona Serratì, Alessandra Modesti, Mario Del Rosso, Gabriella Fibbi

Research output: Contribution to journalArticle

Abstract

Endothelial cell caveolar-rafts are considered functional platforms that recruit several pro-angiogenic molecules to realize an efficient angiogenic program. Here we studied the differential caveolar-raft protein composition of endothelial colony-forming cells following stimulation with VEGF, which localizes in caveolae on interaction with its type-2 receptor. Endothelial colony-forming cells are a cell population identified in human umbilical blood that show all the properties of an endothelial progenitor cell and a high proliferative rate. Two-dimensional gel electrophoresis analysis was coupled with mass spectrometry to identify candidate proteins. The twenty-eight differentially expressed protein spots were grouped according to their function using Gene Ontology classification. In particular, functional categories relative to cell death inhibition and hydrogen peroxide metabolic processes resulted enriched. In these categories, Peroxiredoxin-2 and 6, that control hydrogen peroxide metabolic processes, are the main enriched molecules together with the anti-apoptotic 78 kDa glucose regulated protein. Some of the proteins we identified had never before identified as caveolar-raft components. Other identified proteins include calpain small sub-unit-1, known to mediates angiogenic response to VEGF, gelsolin, which regulates stress fiber assembly, and annexin A3, an angiogenic mediator that induces VEGF production. We validated the functional activity of the above proteins, showing that the siRNA silencing of these resulted in the inhibition of capillary morphogenesis. Overall, our data show that VEGF stimulation triggers the caveolar-raft recruitment of proteins that warrant a physiological amount of reactive oxygen species to maintain a proper angiogenic function of endothelial colony-forming cells and preserve the integrity of the actin cytoskeleton.

Original languageEnglish
Pages (from-to)1926-1938
Number of pages13
JournalMolecular and Cellular Proteomics
Volume12
Issue number7
DOIs
Publication statusPublished - 2013

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry
  • Medicine(all)

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    Chillà, A., Magherini, F., Margheri, F., Laurenzana, A., Gamberi, T., Bini, L., Bianchi, L., Danza, G., Mazzanti, B., Serratì, S., Modesti, A., Del Rosso, M., & Fibbi, G. (2013). Proteomic identification of VEGF-dependent protein enrichment to membrane caveolar-raft microdomains in endothelial progenitor cells. Molecular and Cellular Proteomics, 12(7), 1926-1938. https://doi.org/10.1074/mcp.M112.024638