Basement-membrane heparan sulfate proteoglycan binds to laminin by its heparan sulfate chains and to nidogen by sites in the protein core.
A large, low-density form of heparan sulfate proteoglycan was isolated from the Engelbreth-Holm-Swarm (EHS) tumor and demonstrated to bind in immobilized-ligand assays to laminin fragment E3, collagen type IV, fibronectin and nidogen. The first three ligands mainly recognize the heparan sulfate chains, as shown by inhibition with heparin and heparan sulfate and by the failure to bind to the proteoglycan protein core. Nidogen, obtained from the EHS tumor or in recombinant form, binds exclusively to the protein core in a heparin-insensitive manner. Studies with other laminin fragments indicate that the fragment E3 possesses a unique binding site of laminin for the proteoglycan. A major binding site of nidogen was localized to its central globular domain G2 by using overlapping fragments. This allows for the formation of ternary complexes between laminin, nidogen and proteoglycan, suggesting a key role for nidogen in basement-membrane assembly. Evidence is provided for a second proteoglycan-binding site in the C-terminal globule G3 of nidogen, but this interaction prevents the formation of such ternary complexes. Therefore, the G3-mediated nidogen binding to laminin and proteoglycan are mutually exclusive.
Battaglia
C
C
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
Mayer
U
U
Aumailley
M
M
Timpl
R
R
D001485:Basement Membrane / Q000737:chemistry*; D001665:Binding Sites; D003094:Collagen / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D006493:Heparin / Q000031:analogs & derivatives* / Q000378:metabolism; D007797:Laminin / Q000378:metabolism*; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009374:Neoplasms, Experimental / Q000737:chemistry; D010446:Peptide Fragments / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*
D005353:Fibronectins; D007797:Laminin; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011509:Proteoglycans; C036090:heparin proteoglycan; C029967:nidogen; D006493:Heparin; D003094:Collagen