The 140-kilodalton antiangiogenic fragment of thrombospondin-1 binds to basic fibroblast growth factor

Giulia Taraboletti, Dorina Belotti, Patrizia Borsotti, Veronica Vergani, Marco Rusnati, Marco Presta, Raffaella Giavazzi

Research output: Contribution to journalArticlepeer-review

Abstract

Thrombospondin-1 (TSP) inhibits the angiogenic activity of basic fibroblast growth factor (bFGF). Here we address the hypothesis of a direct interaction between TSP and bFGF. Gel permeation chromatography and cross- linking experiments demonstrated that bFGF binds to TSP in solution. bFGF also bound to immobilized TSP in a solid-phase assay. Binding was dose- dependent, with a K(d) in the nanomolar range, and was inhibited by anti-TSP antibodies. The 140-kDa carboxyl-terminal fragment of TSP, but not the 25- kDa heparin-binding fragment, fully retained the bFGF binding capacity. Accordingly, binding was inhibited by monoclonal antibodies directed against this fragment. Heparin completely blocked bFGF binding to TSP and to the 140- kDa fragment. TSP and its 140-kDa fragment inhibited the binding of bFGF to endothelial cells at concentrations (≤ 100 nM) that inhibited endothelial cell proliferation but not motility. Low-affinity binding was inhibited more than high-affinity binding (up to 76 and 41% inhibition, respectively), and the inhibition was reversed by anti-TSP antibodies. Vitronectin and transforming growth factor β, potentially associated with TSP, did not affect bFGF binding to endothelial cells. Although TSP did not affect the activation of the high-affinity receptors, it reduced the long-term internalization of bFGF. We conclude that TSP binds to bFGF through a domain within its 140-kDa fragment, a mechanism that might affect bFGF interaction with endothelial cells, activity, and association with the extracellular matrix.

Original languageEnglish
Pages (from-to)471-479
Number of pages9
JournalCell Growth and Differentiation
Volume8
Issue number4
Publication statusPublished - Apr 1997

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Fingerprint Dive into the research topics of 'The 140-kilodalton antiangiogenic fragment of thrombospondin-1 binds to basic fibroblast growth factor'. Together they form a unique fingerprint.

Cite this