HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 280, Issue 44, 37217-37224, November 4, 2005

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 280/44/37217    most recent M508060200v3 M508060200v2 M508060200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Gingras, A. R. Articles by Emsley, J. Search for Related Content PubMed PubMed Citation Articles by Gingras, A. R. Articles by Emsley, J. Social Bookmarking                      What's this?

Mapping and Consensus Sequence Identification for Multiple Vinculin Binding Sites within the Talin Rod^*

Alexandre R. Gingras1, Wolfgang H. Ziegler1, Ronald Frank¶, Igor L. Barsukov, Gordon C. K. Roberts, David R. Critchley, and Jonas Emsley||2

From the Department of Biochemistry, University of Leicester, Leicester LE1 7RH, United Kingdom, the Interdisciplinary Centre for Clinical Research (IZKF) Leipzig, Faculty of Medicine, University of Leipzig, D-04103 Leipzig, Germany, the ^¶Department of Chemical Biology, German Research Centre for Biotechnology, D-38124 Braunschweig, Germany, and the ^||Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, Nottingham NG72RD, United Kingdom

The interaction between the cytoskeletal proteins talin and vinculin plays a key role in integrin-mediated cell adhesion and migration. Three vinculin binding sites (VBS1-3) have previously been identified in the talin rod using a yeast two-hybrid assay. To extend these studies, we spot-synthesized a series of peptides spanning all the -helical regions predicted for the talin rod and identified eight additional VBSs, two of which overlap key functional regions of the rod, including the integrin binding site and C-terminal actin binding site. The talin VBS -helices bind to a hydrophobic cleft in the N-terminal vinculin Vd1 domain. We have defined the specificity of this interaction by spot-synthesizing a series of 25-mer talin VBS1 peptides containing substitutions with all the commonly occurring amino acids. The consensus for recognition is LXXAAXXVAXX- VXXLIXXA with distinct classes of hydrophobic side chains at positions 1, 4, 5, 8, 9, 12, 15, and 16 required for vinculin binding. Positions 1, 8, 12, 15, and 16 require an aliphatic residue and will not tolerate alanine, whereas positions 4, 5, and 9 are less restrictive. These preferences are common to all 11 VBS sequences with a minor variation occurring in one case. A crystal structure of this variant VBS peptide in complex with the vinculin Vd1 domain reveals a subtly different mode of vinculin binding.

Received for publication, July 22, 2005

The atomic coordinates and structure factors (code 1ZW2 and 1ZW3) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* This work was funded by grants from Biotechnology and Biological Sciences and Research Council, the Wellcome Trust, and the German Research Council. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. S1.

¹ Both authors contributed equally to this work.

² To whom correspondence should be addressed. Tel.: 44-1-158-467-092; Fax: 44-1-158-468-002; E-mail: jonas.emsley{at}nottingham.ac.uk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				B. T. Goult, N. Bate, N. J. Anthis, K. L. Wegener, A. R. Gingras, B. Patel, I. L. Barsukov, I. D. Campbell, G. C. K. Roberts, and D. R. Critchley The Structure of an Interdomain Complex That Regulates Talin Activity J. Biol. Chem., May 29, 2009; 284(22): 15097 - 15106. [Abstract] [Full Text] [PDF]

				M. Himmel, A. Ritter, S. Rothemund, B. V. Pauling, K. Rottner, A. R. Gingras, and W. H. Ziegler Control of High Affinity Interactions in the Talin C Terminus: HOW TALIN DOMAINS COORDINATE PROTEIN DYNAMICS IN CELL ADHESIONS J. Biol. Chem., May 15, 2009; 284(20): 13832 - 13842. [Abstract] [Full Text] [PDF]

				A. R. Gingras, W. H. Ziegler, A. A. Bobkov, M. G. Joyce, D. Fasci, M. Himmel, S. Rothemund, A. Ritter, J. G. Grossmann, B. Patel, et al. Structural Determinants of Integrin Binding to the Talin Rod J. Biol. Chem., March 27, 2009; 284(13): 8866 - 8876. [Abstract] [Full Text] [PDF]

				K. R. Legate, S. A. Wickstrom, and R. Fassler Genetic and cell biological analysis of integrin outside-in signaling Genes & Dev., February 15, 2009; 23(4): 397 - 418. [Abstract] [Full Text] [PDF]

				A. del Rio, R. Perez-Jimenez, R. Liu, P. Roca-Cusachs, J. M. Fernandez, and M. P. Sheetz Stretching Single Talin Rod Molecules Activates Vinculin Binding Science, January 30, 2009; 323(5914): 638 - 641. [Abstract] [Full Text] [PDF]

				K. R. Legate and R. Fassler Mechanisms that regulate adaptor binding to {beta}-integrin cytoplasmic tails J. Cell Sci., January 15, 2009; 122(2): 187 - 198. [Abstract] [Full Text] [PDF]

				S. Rodius, O. Chaloin, M. Moes, E. Schaffner-Reckinger, I. Landrieu, G. Lippens, M. Lin, J. Zhang, and N. Kieffer The Talin Rod IBS2 {alpha}-Helix Interacts with the {beta}3 Integrin Cytoplasmic Tail Membrane-proximal Helix by Establishing Charge Complementary Salt Bridges J. Biol. Chem., August 29, 2008; 283(35): 24212 - 24223. [Abstract] [Full Text] [PDF]

				D. R. Critchley and A. R. Gingras Talin at a glance J. Cell Sci., May 1, 2008; 121(9): 1345 - 1347. [Full Text] [PDF]

				C. Le Clainche and M.-F. Carlier Regulation of Actin Assembly Associated With Protrusion and Adhesion in Cell Migration Physiol Rev, April 1, 2008; 88(2): 489 - 513. [Abstract] [Full Text] [PDF]

				M. Moes, S. Rodius, S. J. Coleman, S. J. Monkley, E. Goormaghtigh, L. Tremuth, C. Kox, P. P. G. van der Holst, D. R. Critchley, and N. Kieffer The Integrin Binding Site 2 (IBS2) in the Talin Rod Domain Is Essential for Linking Integrin beta Subunits to the Cytoskeleton J. Biol. Chem., June 8, 2007; 282(23): 17280 - 17288. [Abstract] [Full Text] [PDF]

				H. Chen, D. M. Choudhury, and S. W. Craig Coincidence of Actin Filaments and Talin Is Required to Activate Vinculin J. Biol. Chem., December 29, 2006; 281(52): 40389 - 40398. [Abstract] [Full Text] [PDF]

				D. M. Cohen, B. Kutscher, H. Chen, D. B. Murphy, and S. W. Craig A Conformational Switch in Vinculin Drives Formation and Dynamics of a Talin-Vinculin Complex at Focal Adhesions J. Biol. Chem., June 9, 2006; 281(23): 16006 - 16015. [Abstract] [Full Text] [PDF]

				B. Patel, A. R. Gingras, A. A. Bobkov, L. M. Fujimoto, M. Zhang, R. C. Liddington, D. Mazzeo, J. Emsley, G. C. K. Roberts, I. L. Barsukov, et al. The Activity of the Vinculin Binding Sites in Talin Is Influenced by the Stability of the Helical Bundles That Make Up The Talin Rod J. Biol. Chem., March 17, 2006; 281(11): 7458 - 7467. [Abstract] [Full Text] [PDF]

				P. R. J. Bois, B. P. O'Hara, D. Nietlispach, J. Kirkpatrick, and T. Izard The Vinculin Binding Sites of Talin and {alpha}-Actinin Are Sufficient to Activate Vinculin J. Biol. Chem., March 17, 2006; 281(11): 7228 - 7236. [Abstract] [Full Text] [PDF]