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The intermediate filament protein vimentin binds specifically to a recombinant integrin α2/β1 cytoplasmic tail complex and co-localizes with native α2/β1 in endothelial cell focal adhesions
Kreis, Stephanie; Schönfeld, H.-J. B; Melchior, C. A et al.
2005In Experimental Cell Research, 305 (1), p. 110-121
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Keywords :
Cytoplasmic tail; Integrin α2β1; Interaction; Intermediate filament; Recombinant complex; Vimentin
Abstract :
[en] Integrin receptors are crucial players in cell adhesion and migration. Identification and characterization of cellular proteins that interact with their short α and β cytoplasmic tails will help to elucidate the molecular mechanisms by which integrins mediate bi-directional signaling across the plasma membrane. Integrin α2β1 is a major collagen receptor but to date, only few proteins have been shown to interact with the α2 cytoplasmic tail or with the α2β1 complex. In order to identify novel binding partners of a α2β1cytoplasmic 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-α2 and GST-Jun α2 bound His-tagged calreticulin while GST-β1 and GST-Fos β1 proteins bound talin. In screening assays for novel binding partners, the immobilized GST-Jun α2/GST-Fos β1 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 αvβ3-positive focal contacts. Here, we provide evidence that this interaction also occurs with α2β1-enriched focal adhesions and we further show that this association is lost after prolonged adhesion of endothelial cells to collagen. © 2005 Elsevier Inc. All rights reserved.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Kreis, Stephanie ;  University of Luxembourg > Faculty of Science, Technology and Communication (FSTC) > Life Science Research Unit
Schönfeld, H.-J. B;  F. Hoffmann-La Roche Ltd., Pharmaceuticals Division, Preclin. Res., Vasc. and Metab. Dis., CH-4070 Basel, Switzerland
Melchior, C. A;  ACTELION Pharmaceuticals Ltd., Innovation Center, Gewerbestrasse 16, CH-4123 Allschwil, Switzerland
Steiner, B. B C
Kieffer, N. A
Language :
English
Title :
The intermediate filament protein vimentin binds specifically to a recombinant integrin α2/β1 cytoplasmic tail complex and co-localizes with native α2/β1 in endothelial cell focal adhesions
Publication date :
2005
Journal title :
Experimental Cell Research
ISSN :
1090-2422
Publisher :
Academic Press, Orlando, United States - Florida
Volume :
305
Issue :
1
Pages :
110-121
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
cited By (since 1996)36 Scopus
Available on ORBilu :
since 19 August 2013

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