A novel synthetic peptide from the B1 chain of laminin with heparin-binding and cell adhesion-promoting activities.
Recent studies using solid-phase-binding assays and electron microscopy suggested the presence of a heparin-binding domain between the inner globule of a lateral short arm and the cross region of laminin. Using the information from the amino acid sequence of the B1 chain of laminin, several peptides were synthesized from areas with a low hydropathy index and a high density of lysines and/or arginines. One of these, peptide F-9 (RYVVLPRPVCFEKGMNYTVR), which is derived from the inner globular domain of the lateral short arm, demonstrated specific binding to heparin. This was tested in direct solid-phase binding assays by coating the peptide either on nitrocellulose or on polystyrene and in indirect competition assays where the peptide was in solution and either laminin or heparin was immobilized on a solid support. The binding of [3H]heparin to peptide F-9 was dramatically reduced when heparin but not other glycosaminoglycans other than heparin (dextran sulfate, dermatan sulfate) were used in competition assays. Modification of the free amino groups of peptide F-9 by acetylation abolished its ability to inhibit the binding of [3H]heparin to laminin on polystyrene surfaces. Peptide F-9 promoted the adhesion of various cell lines (melanoma, fibrosarcoma, glioma, pheochromocytoma) and of aortic endothelial cells. Furthermore, when peptide F-9 was present in solution, it inhibited the adhesion of melanoma cells to laminin-coated substrates. These findings suggest that peptide F-9 defines a novel heparin-binding and cell adhesion-promoting site on laminin.
Charonis
A S
AS
Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis 55455.
Skubitz
A P
AP
Koliakos
G G
GG
Reger
L A
LA
Dege
J
J
Vogel
A M
AM
Wohlhueter
R
R
Furcht
L T
LT
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002448:Cell Adhesion; D002460:Cell Line; D005354:Fibrosarcoma; D005910:Glioma; D006493:Heparin / Q000378:metabolism*; D007797:Laminin / Q000378:metabolism*; D008545:Melanoma; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism*; D010673:Pheochromocytoma; D014407:Tumor Cells, Cultured
D007797:Laminin; D010446:Peptide Fragments; D006493:Heparin