An Arg-Gly-Asp sequence within thrombin promotes endothelial cell adhesion

doi:10.1083/jcb.112.2.335

This Article

	Full Text (PDF, 2817K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Bar-Shavit, R.
	Articles by Dejana, E.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Bar-Shavit, R.
	Articles by Dejana, E.


Social Bookmarking

	What's this?

ARTICLES

An Arg-Gly-Asp sequence within thrombin promotes endothelial cell adhesion

R Bar-Shavit, V Sabbah, MG Lampugnani, PC Marchisio, JW Fenton 2d, I Vlodavsky and E Dejana
Department of Oncology, Hadassah University Hospital, Jerusalem, Israel.

Thrombin, in addition to its central role in hemostasis, possesses diverse cellular bioregulatory functions implicated in wound healing, inflammation, and atherosclerosis. In the present study we demonstrate that thrombin molecules modified either at the procoagulant or catalytic sites induce endothelial cell (EC) adhesion, spreading, and cytoskeletal reorganization. The most potent adhesive thrombin analogue (NO2-alpha-thrombin) was obtained by nitration of tyrosine residues. The cell adhesion promoting activity of NO2-alpha-thrombin was blocked upon the formation of thrombin-antithrombin III (ATIII) complexes and by antiprothrombin antibodies, but was unaffected by hirudin. Arg-Gly- Asp-containing peptides, fully inhibited EC adhesion to NO2-alpha- thrombin, while synthetic peptides corresponding to thrombin "Loop B" mitogenic site and the thrombin-derived chemotactic fragment "CB67- 129", were uneffective. Immunofluorescence studies indicated that EC adhesion to NO2-alpha-thrombin was followed by cell spreading, actin microfilament assembly, and formation of focal contacts. By the use of specific antibodies, the vitronectin (vn) receptor (alpha v beta 3) was found to be localized in clusters upon cell adhesion to NO2-alpha- thrombin. An anti alpha v beta 3 antibody blocked EC adhesion and spreading while antifibronectin (fn) receptor (alpha 5 beta 1) antibodies were uneffective. While native thrombin exhibited a very low cell attachment activity, thrombin that was incubated at 37 degrees C before coating of plastic surfaces induced EC attachment and spreading. We propose that under certain conditions the naturally hindered RGD domain within thrombin is exposed for interaction with alpha v beta 3 on EC. This in turn promotes cell adhesion, spreading, and reorganization of cytoskeletal elements, which may altogether contribute to repair mechanisms in the disturbed vessel wall. This study defines a new biological role of thrombin and characterizes a new recognition mechanism on EC for this molecule.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Waldeck-Weiermair, M., Zoratti, C., Osibow, K., Balenga, N., Goessnitzer, E., Waldhoer, M., Malli, R., Graier, W. F. (2008). Integrin clustering enables anandamide-induced Ca2+ signaling in endothelial cells via GPR55 by protection against CB1-receptor-triggered repression. J. Cell Sci. 121: 1704-1717 [Abstract] [Full Text]
Stefansson, S., Su, E. J., Ishigami, S., Cale, J. M., Gao, Y., Gorlatova, N., Lawrence, D. A. (2007). The Contributions of Integrin Affinity and Integrin-Cytoskeletal Engagement in Endothelial and Smooth Muscle Cell Adhesion to Vitronectin. J. Biol. Chem. 282: 15679-15689 [Abstract] [Full Text]
Polenghi, A., Bossi, F., Fischetti, F., Durigutto, P., Cabrelle, A., Tamassia, N., Cassatella, M. A., Montecucco, C., Tedesco, F., de Bernard, M. (2007). The Neutrophil-Activating Protein of Helicobacter pylori Crosses Endothelia to Promote Neutrophil Adhesion In Vivo. J. Immunol. 178: 1312-1320 [Abstract] [Full Text]
Lechner, A. M., Assfalg-Machleidt, I., Zahler, S., Stoeckelhuber, M., Machleidt, W., Jochum, M., Nagler, D. K. (2006). RGD-dependent Binding of Procathepsin X to Integrin {alpha}vbeta3 Mediates Cell-adhesive Properties. J. Biol. Chem. 281: 39588-39597 [Abstract] [Full Text]
Papaconstantinou, M. E., Carrell, C. J., Pineda, A. O., Bobofchak, K. M., Mathews, F. S., Flordellis, C. S., Maragoudakis, M. E., Tsopanoglou, N. E., Di Cera, E. (2005). Thrombin Functions through Its RGD Sequence in a Non-canonical Conformation. J. Biol. Chem. 280: 29393-29396 [Abstract] [Full Text]
Stouffer, G.A., Smyth, S.S. (2003). Effects of Thrombin on Interactions Between {beta}3-Integrins and Extracellular Matrix in Platelets and Vascular Cells. Arterioscler. Thromb. Vasc. Bio. 23: 1971-1978 [Abstract] [Full Text]
Seymour, M. L., Zaidi, N. F., Hollenberg, M. D., MacNaughton, W. K. (2003). PAR1-dependent and independent increases in COX-2 and PGE2 in human colonic myofibroblasts stimulated by thrombin. Am. J. Physiol. Cell Physiol. 284: C1185-C1192 [Abstract] [Full Text]
Tsopanoglou, N. E., Andriopoulou, P., Maragoudakis, M. E. (2002). On the mechanism of thrombin-induced angiogenesis: involvement of alpha vbeta 3-integrin. Am. J. Physiol. Cell Physiol. 283: C1501-C1510 [Abstract] [Full Text]
Allain, F., Vanpouille, C., Carpentier, M., Slomianny, M.-C., Durieux, S., Spik, G. (2002). Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix. Proc. Natl. Acad. Sci. USA 10.1073/pnas.052284899v1 [Abstract] [Full Text]
Dallabrida, S. M., Falls, L. A., Farrell, D. H. (2000). Factor XIIIa supports microvascular endothelial cell adhesion and inhibits capillary tube formation in fibrin. Blood 95: 2586-2592 [Abstract] [Full Text]
Cal, S., Freije, J. M.P., López, J. M., Takada, Y., López-Otín, C. (2000). ADAM 23/MDC3, a Human Disintegrin That Promotes Cell Adhesion via Interaction with the alpha vbeta 3 Integrin through an RGD-independent Mechanism. Mol. Biol. Cell 11: 1457-1469 [Abstract] [Full Text]
Byzova, T. V., Plow, E. F. (1998). Activation of {alpha}V{beta}3 on Vascular Cells Controls Recognition of Prothrombin. JCB 143: 2081-2092 [Abstract] [Full Text]
Rusnati, M., Tanghetti, E., Dell'Era, P., Gualandris, A., Presta, M. (1997). alpha vbeta 3 Integrin Mediates the Cell-adhesive Capacity and Biological Activity of Basic Fibroblast Growth Factor (FGF-2) in Cultured Endothelial Cells. Mol. Biol. Cell 8: 2449-2461 [Abstract] [Full Text]
Ueki, S, Takagi, J, Saito, Y (1996). Dual functions of transglutaminase in novel cell adhesion. J. Cell Sci. 109: 2727-2735 [Abstract]
Even-Ram, S. C., Maoz, M., Pokroy, E., Reich, R., Katz, B.-Z., Gutwein, P., Altevogt, P., Bar-Shavit, R. (2001). Tumor Cell Invasion Is Promoted by Activation of Protease Activated Receptor-1 in Cooperation with the alpha vbeta 5 Integrin. J. Biol. Chem. 276: 10952-10962 [Abstract] [Full Text]
Allain, F., Vanpouille, C., Carpentier, M., Slomianny, M.-C., Durieux, S., Spik, G. (2002). Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix. Proc. Natl. Acad. Sci. USA 99: 2714-2719 [Abstract] [Full Text]