Cooperation between thrombospondin-1 type 1 repeat peptides and alpha(v)beta(3) integrin ligands to promote melanoma cell spreading and focal adhesion kinase phosphorylation.
CD47-binding sequences from the carboxyl-terminal domain of thrombospondin-1 (TSP1) are known to regulate activity of the alpha(v)beta(3) integrin (Gao, G., Lindberg, F. P., Dimitry, J. M., Brown, E. J., and Frazier, W. A. (1996) J. Cell Biol. 135, 533-544). Here we show that peptides from the type 1 repeats of TSP1 also stimulate alpha(v)beta(3) integrin function in melanoma cells. Addition of soluble peptide 246 (KRFKQDGGWSHWSPWSS) enhances spreading of A2058 melanoma cells on several alpha(v)beta(3) integrin ligands, including vitronectin, recombinant TSP1 fragments containing the Arg-Gly-Asp sequence, and native TSP1. This activity requires the Trp residues and is independent of CD36-binding sequences in the type 1 repeats. Recombinant type 1 repeats expressed as a glutathione S-transferase fusion protein also enhance spreading on vitronectin and TSP1. Activation of alpha(v)beta(3) integrin by the soluble peptide 246 stimulates organization of F-actin and increases tyrosine phosphorylation of focal adhesion kinase. In contrast, direct adhesion of melanoma cells on immobilized peptide 246 inhibits tyrosine phosphorylation of focal adhesion kinase. Stimulation of alpha(v)beta(3) integrin function by the type 1 repeat peptide differs from that induced by CD47-binding TSP1 peptides in that heparan sulfate proteoglycans are required and pertussis toxin does not inhibit the former activity. Thus, the type 1 repeats contain a second sequence of TSP1 that can enhance alpha(v)beta(3) integrin signaling, and these two sequences stimulate recognition of both vitronectin and TSP1 by the alpha(v)beta(3) integrin.
Sipes
J M
JM
Laboratory of Pathology, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Krutzsch
H C
HC
Lawler
J
J
Roberts
D D
DD
D000595:Amino Acid Sequence; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D048429:Cell Size / Q000187:drug effects; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019812:Heparan Sulfate Proteoglycans / Q000494:pharmacology; D006801:Humans; D008024:Ligands; D008546:Melanoma, Experimental / Q000503:physiopathology*; D008954:Models, Biological; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010446:Peptide Fragments / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000378:metabolism*; D020449:Repetitive Sequences, Amino Acid; D015398:Signal Transduction; D019700:Thrombospondin 1 / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D019096:Vitronectin / Q000494:pharmacology
D015815:Cell Adhesion Molecules; D019812:Heparan Sulfate Proteoglycans; D008024:Ligands; D009842:Oligopeptides; D010446:Peptide Fragments; D019030:Receptors, Vitronectin; D019700:Thrombospondin 1; D019096:Vitronectin; C047981:arginyl-glycyl-aspartic acid; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human