An angiogenic laminin site and its antagonist bind through the alpha(v)beta3 and alpha5beta1 integrins.
Angiogenesis is important for wound healing, tumor growth, and metastasis. Endothelial cells differentiate into capillary-like structures on a laminin-1-rich matrix (Matrigel). We previously identified 20 angiogenic sites on laminin-1 (alpha1beta1gamma1) by screening 559 overlapping synthetic peptides. C16, the most potent gamma1 chain peptide, blocked laminin-1-mediated adhesion and was the only gamma1 chain peptide to block attachment to both collagen I and fibronectin. This suggested that C16 was acting via a receptor common to these substrates. We demonstrated that C16 is angiogenic in vivo. Affinity chromatography identified the integrins alpha5beta1 and alpha(v)beta3 as surface receptors. Blocking antibodies confirmed the role of these receptors in C16 adhesion. C16 does not contain an RGD sequence and, as expected, an RGD-containing peptide did not block C16 adhesion nor did C16 act via MAP kinase phosphorylation. Furthermore, we identified a C16 scrambled sequence, C16S, which antagonizes the angiogenic activity of bFGF and of C16 by binding to the same receptors. Because the laminin gamma1 chain is ubiquitous in most tissues, C16 is likely an important functional site. Since the biological activity of C16 is blocked by a scrambled peptide, C16S may serve as an anti-angiogenic therapeutic agent.
Ponce
M L
ML
Craniofacial Developmental Biology and Regeneration Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
Nomizu
M
M
Kleinman
H K
HK
D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000254:growth & development; D001665:Binding Sites; D002448:Cell Adhesion; D002642:Chick Embryo; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors; D039121:Integrin alpha6beta1; D016023:Integrins / Q000276:immunology / Q000378:metabolism*; D007797:Laminin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017474:Receptors, Laminin / Q000276:immunology / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000276:immunology / Q000378:metabolism*
D039121:Integrin alpha6beta1; D016023:Integrins; D007797:Laminin; D010446:Peptide Fragments; D017474:Receptors, Laminin; D019030:Receptors, Vitronectin; C121346:laminin 1; D016222:Fibroblast Growth Factor 2; D020928:Mitogen-Activated Protein Kinases