Urokinase binding to laminin-nidogen. Structural requirements and interactions with heparin.
Recently we have shown that heparin and related sulfated polyanions are low-affinity ligands of the kringle domain in the amino-terminal region (ATF) of human urokinase (u-PA), and proposed that this may facilitate loading of u-PA onto its receptor at the focal contacts between adherent cells and their matrix. We have now tested other components of the cell matrix (fibronectin, vitronectin, thrombospondin and laminin-nidogen) for u-PA binding, and found that laminin-nidogen is also a ligand of the u-PA ATF. Direct binding assays and competition binding assays with defined fragments of laminin-nidogen showed that there are u-PA binding sites in fragment E4 of laminin as well as in nidogen. The long-arm terminal domain of laminin (fragment E3), which contains a heparin-binding site, competed for binding of u-PA to immobilised heparin. However nidogen, which does not bind to heparin, also inhibited binding of u-PA to heparin, and this effect was also observed with recombinant nidogen and with a fragment of nidogen lacking the carboxy-terminal domain. Direct binding assays confirmed that u-PA binds to nidogen through a site in the u-PA ATF. We conclude that u-PA binds to laminin-nidogen by interactions involving the ATF region of u-PA, the E4 domain of laminin and the rod or amino-terminal regions of nidogen. Since nidogen is suggested to be an important bridging molecule in the maintenance of the supramolecular organization in basement membranes, the presence of a binding site for u-PA in nidogen indicates a role for plasminogen activation in basement membrane remodelling.
Stephens
R W
RW
Department of Virology, University of Helsinki, Finland.
Aumailley
M
M
Timpl
R
R
Reisberg
T
T
Tapiovaara
H
H
Myöhänen
H
H
Murphy-Ullrich
J
J
Vaheri
A
A
D001667:Binding, Competitive; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D007797:Laminin; D008562:Membrane Glycoproteins; C029967:nidogen; D006493:Heparin; D014568:Urokinase-Type Plasminogen Activator