title: Proliferation and migration activities of fibroblast growth factor-2 in endothelial cells are modulated by its direct interaction with heparin affin regulatory peptide.
abstract: Angiogenesis is the physiological process involving the growth of new blood vessels from pre-existing vessels. In normal or pathological angiogenesis, angiogenic growth factors activate cognate receptors on endothelial cells. Fibroblast growth factor-2 (FGF-2) and heparin affin regulatory peptide (HARP) are two heparin-binding growth factors and were described for their pro-angiogenic properties on human umbilical endothelial cells (HUVEC). We now show that HARP acts as a mediator of FGF-2's stimulatory effects, since it is able to inhibit the proliferation and migration of HUVEC induced by FGF-2. We demonstrate by ELISA and optical biosensor binding assay that HARP and FGF-2 interact through direct binding. We have adapted a previously developed structural proteomics method for the identification of residues involved in protein-protein interactions. Application of this method showed that two sequences in HARP were involved in binding FGF-2. One was in the C-thrombospondin type 1 repeat (C-TSR-1) domain and the other in the C-terminal domain of HARP. The identification of these regions as mediating the binding of FGF-2 was confirmed by ELISA using synthetic peptides, which are as well mediators of FGF-2-induced proliferation, migration and tubes formation on HUVEC in vitro. These results imply that besides a regulation of the proliferation, migration and angiogenesis of HUVEC by direct interaction of FGF-2 with its receptors, an alternative pathway exists involving its binding to growth factors such as HARP.
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France. Electronic address: ccliadossantos@gmail.com.
authors.forename: Célia
authors.initials: C
authors.lastname: Dos Santos
authors.affiliation: IMRB INSERM, U955, Equipe 07, Faculté de Médecine, 8 rue du Général Sarrail, 94010 Créteil, France.
authors.forename: Charly
authors.initials: C
authors.lastname: Blanc
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France.
authors.forename: Rania
authors.initials: R
authors.lastname: Elahouel
authors.affiliation: Department of Structural and Chemical Biology, Institute of Integrative Biology, Biosciences Building, University of Liverpool, Liverpool L69 7ZB, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Carpentier
authors.affiliation: Department of Structural and Chemical Biology, Institute of Integrative Biology, Biosciences Building, University of Liverpool, Liverpool L69 7ZB, UK.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ori
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France.
authors.forename: José
authors.initials: J
authors.lastname: Courty
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France.
authors.forename: Yamina
authors.initials: Y
authors.lastname: Hamma-Kourbali
authors.affiliation: Department of Structural and Chemical Biology, Institute of Integrative Biology, Biosciences Building, University of Liverpool, Liverpool L69 7ZB, UK.
authors.forename: David G
authors.initials: DG
authors.lastname: Fernig
authors.affiliation: Laboratoire CRRET, CNRS, Université Paris Est, Avenue du Général de Gaulle, 94010 Créteil Cedex, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Delbé
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D002352:Carrier Proteins / Q000378:metabolism* / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism* / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism* / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D016207:Cytokines; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; C066845:pleiotrophin
keywords: Growth factors; Interaction; Migration; Proliferation; Protect and label