Urokinase regulates vitronectin binding by controlling urokinase receptor oligomerization

Nicolai Sidenius, Annapaola Andolfo, Riccardo Fesce, Francesco Blasi

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Adhesion of monocytes to the extracellular matrix is mediated by a direct high affinity interaction between cell-surface urokinase-type plasminogen activator (uPA) receptor (uPAR) and the extracellular matrix protein vitronectin. We demonstrate a tight connection between uPA-regulated uPAR oligomerization and high affinity binding to immobilized vitronectin. We find that binding of soluble uPAR (suPAR) to immobilized vitronectin is strictly ligand-dependent with a linear relationship between the observed binding and the concentration of ligand added. Nevertheless, a comparison of experimentally obtained binding curves to those generated using a simple equilibrium model suggests that the high affinity vitronectin-binding pro-uPA-suPAR complex contains two molecules of suPAR. In co-immunoprecipitation experiments, using different epitopetagged suPAR molecules, suPAR/suPAR co-immunoprecipitation displayed a similar uPA dose dependence as that observed for vitronectin binding, demonstrating that the high affinity vitronectin-binding complex indeed contains oligomeric suPAR. Structurally, the kringle domain of uPA was found to be critical for the formation of the vitronectin-binding competent complex because the amino-terminal fragment, but not the growth factor-like domain, behaved as a full-length uPA. Our data represent the first demonstration of functional, ligand-induced uPAR oligomerization having extensive implications for glycosylphosphatidylinositolanchored receptors in general, and for the biology of the uPA/uPAR system in particular.

Original languageEnglish
Pages (from-to)27982-27990
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number31
DOIs
Publication statusPublished - Aug 2 2002

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Vitronectin
Oligomerization
Urokinase-Type Plasminogen Activator
Urokinase Plasminogen Activator Receptors
saruplase
Ligands
Immunoprecipitation
Kringles
Molecules
Extracellular Matrix Proteins
Cell Communication
Extracellular Matrix
Monocytes
Intercellular Signaling Peptides and Proteins
Demonstrations
Adhesion

ASJC Scopus subject areas

  • Biochemistry

Cite this

Urokinase regulates vitronectin binding by controlling urokinase receptor oligomerization. / Sidenius, Nicolai; Andolfo, Annapaola; Fesce, Riccardo; Blasi, Francesco.

In: Journal of Biological Chemistry, Vol. 277, No. 31, 02.08.2002, p. 27982-27990.

Research output: Contribution to journalArticle

Sidenius, Nicolai ; Andolfo, Annapaola ; Fesce, Riccardo ; Blasi, Francesco. / Urokinase regulates vitronectin binding by controlling urokinase receptor oligomerization. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 31. pp. 27982-27990.
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