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PMID: 1988465 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

The Journal of cell biology ·Vol. 112 ·No. 2 ·1991-01-00 ·Pages 335-44

Bar-Shavit R, Sabbah V, Lampugnani MG, Marchisio PC, Fenton JW, Vlodavsky I, Dejana E

Abstract

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.

MeSH Terms
Amino Acid Sequence Animals Aorta Cattle Cell Adhesion Cells, Cultured Endothelium, Vascular/cytology,metabolism Fluorescent Antibody Technique Humans Molecular Sequence Data Oligopeptides/metabolism Thrombin/antagonists & inhibitors,chemistry,metabolism
Chemicals
Oligopeptides arginyl-glycyl-aspartic acid Thrombin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Bar-Shavit R
Department of Oncology, Hadassah University Hospital, Jerusalem, Israel.
Sabbah V
Lampugnani M G
Marchisio P C
Fenton J W
Vlodavsky I
Dejana E
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.112.2.335
Published
1991-01-00
Pages
335-44
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2288825
Subset
IM
Grants
NCI NIH HHS · CA32089 · United States
External Links
PubMed source DOI: 10.1083/jcb.112.2.335 PMC Full Text View journal details
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