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Title: The intermediate filament protein vimentin binds specifically to a recombinant integrin {alpha}2/{beta}1 cytoplasmic tail complex and co-localizes with native {alpha}2/{beta}1 in endothelial cell focal adhesions

Abstract

Integrin receptors are crucial players in cell adhesion and migration. Identification and characterization of cellular proteins that interact with their short {alpha} and {beta} cytoplasmic tails will help to elucidate the molecular mechanisms by which integrins mediate bi-directional signaling across the plasma membrane. Integrin {alpha}2{beta}1 is a major collagen receptor but to date, only few proteins have been shown to interact with the {alpha}2 cytoplasmic tail or with the {alpha}2{beta}1 complex. In order to identify novel binding partners of a {alpha}2{beta}1cytoplasmic domain complex, we have generated recombinant GST-fusion proteins, incorporating the leucine zipper heterodimerization cassettes of Jun and Fos. To ascertain proper functionality of the recombinant proteins, interaction with natural binding partners was tested. GST-{alpha}2 and GST-Jun {alpha}2 bound His-tagged calreticulin while GST-{beta}1 and GST-Fos {beta}1 proteins bound talin. In screening assays for novel binding partners, the immobilized GST-Jun {alpha}2/GST-Fos {beta}1 heterodimeric complex, but not the single subunits, interacted specifically with endothelial cell-derived vimentin. Vimentin, an abundant intermediate filament protein, has previously been shown to co-localize with {alpha}v{beta}3-positive focal contacts. Here, we provide evidence that this interaction also occurs with {alpha}2{beta}1-enriched focal adhesions and we further show that this association is lost after prolonged adhesion of endothelial cells to collagen.

Authors:
 [1];  [2];  [1];  [2];  [1]
  1. LBPI: Laboratoire de Biologie et Physiologie Integree (CNRS/GDRE-ITI), Universite du Luxembourg, 162A, avenue de la Faiencerie, L-1511 Luxembourg (Luxembourg)
  2. F. Hoffmann-La Roche Ltd., Pharmaceuticals Division, Preclinical Research, Vascular and Metabolic Diseases, CH-4070 Basel (Switzerland)
Publication Date:
OSTI Identifier:
20717581
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 305; Journal Issue: 1; Other Information: DOI: 10.1016/j.yexcr.2004.12.023; PII: S0014-4827(04)00737-2; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ADHESION; COLLAGEN; FILAMENTS; LEUCINE; RECEPTORS

Citation Formats

Kreis, Stephanie, Schoenfeld, Hans-Joachim, Melchior, Chantal, Steiner, Beat, and Kieffer, Nelly. The intermediate filament protein vimentin binds specifically to a recombinant integrin {alpha}2/{beta}1 cytoplasmic tail complex and co-localizes with native {alpha}2/{beta}1 in endothelial cell focal adhesions. United States: N. p., 2005. Web. doi:10.1016/j.yexcr.2004.12.023.
Kreis, Stephanie, Schoenfeld, Hans-Joachim, Melchior, Chantal, Steiner, Beat, & Kieffer, Nelly. The intermediate filament protein vimentin binds specifically to a recombinant integrin {alpha}2/{beta}1 cytoplasmic tail complex and co-localizes with native {alpha}2/{beta}1 in endothelial cell focal adhesions. United States. doi:10.1016/j.yexcr.2004.12.023.
Kreis, Stephanie, Schoenfeld, Hans-Joachim, Melchior, Chantal, Steiner, Beat, and Kieffer, Nelly. Fri . "The intermediate filament protein vimentin binds specifically to a recombinant integrin {alpha}2/{beta}1 cytoplasmic tail complex and co-localizes with native {alpha}2/{beta}1 in endothelial cell focal adhesions". United States. doi:10.1016/j.yexcr.2004.12.023.
@article{osti_20717581,
title = {The intermediate filament protein vimentin binds specifically to a recombinant integrin {alpha}2/{beta}1 cytoplasmic tail complex and co-localizes with native {alpha}2/{beta}1 in endothelial cell focal adhesions},
author = {Kreis, Stephanie and Schoenfeld, Hans-Joachim and Melchior, Chantal and Steiner, Beat and Kieffer, Nelly},
abstractNote = {Integrin receptors are crucial players in cell adhesion and migration. Identification and characterization of cellular proteins that interact with their short {alpha} and {beta} cytoplasmic tails will help to elucidate the molecular mechanisms by which integrins mediate bi-directional signaling across the plasma membrane. Integrin {alpha}2{beta}1 is a major collagen receptor but to date, only few proteins have been shown to interact with the {alpha}2 cytoplasmic tail or with the {alpha}2{beta}1 complex. In order to identify novel binding partners of a {alpha}2{beta}1cytoplasmic domain complex, we have generated recombinant GST-fusion proteins, incorporating the leucine zipper heterodimerization cassettes of Jun and Fos. To ascertain proper functionality of the recombinant proteins, interaction with natural binding partners was tested. GST-{alpha}2 and GST-Jun {alpha}2 bound His-tagged calreticulin while GST-{beta}1 and GST-Fos {beta}1 proteins bound talin. In screening assays for novel binding partners, the immobilized GST-Jun {alpha}2/GST-Fos {beta}1 heterodimeric complex, but not the single subunits, interacted specifically with endothelial cell-derived vimentin. Vimentin, an abundant intermediate filament protein, has previously been shown to co-localize with {alpha}v{beta}3-positive focal contacts. Here, we provide evidence that this interaction also occurs with {alpha}2{beta}1-enriched focal adhesions and we further show that this association is lost after prolonged adhesion of endothelial cells to collagen.},
doi = {10.1016/j.yexcr.2004.12.023},
journal = {Experimental Cell Research},
issn = {0014-4827},
number = 1,
volume = 305,
place = {United States},
year = {2005},
month = {4}
}