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First published online 22 December 2004
doi: 10.1242/jcs.01623

Journal of Cell Science 118, 291-300 (2005)
Published by The Company of Biologists 2005
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Research Article

A specific {alpha}5ß1-integrin conformation promotes directional integrin translocation and fibronectin matrix formation

Katherine Clark1, Roumen Pankov1, Mark A. Travis2,*, Janet A. Askari2, A. Paul Mould2, Susan E. Craig2, Peter Newham2,{ddagger}, Kenneth M. Yamada1 and Martin J. Humphries2,§

1 Craniofacial Developmental Biology and Regeneration Branch, NIDCR, NIH, Bethesda, MA 20892, USA
2 Wellcome Trust Centre for Cell-Matrix Research, School of Biological Sciences, University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK

§ Author for correspondence (e-mail: martin.humphries{at}man.ac.uk)

Accepted 14 October 2004

Integrin adhesion receptors are structurally dynamic proteins that adopt a number of functionally relevant conformations. We have produced a conformation-dependent anti-{alpha}5 monoclonal antibody (SNAKA51) that converts {alpha}5ß1 integrin into a ligand-competent form and promotes fibronectin binding. In adherent fibroblasts, SNAKA51 preferentially bound to integrins in fibrillar adhesions. Clustering of integrins expressing this activation epitope induced directional translocation of {alpha}5ß1, mimicking fibrillar adhesion formation. Priming of {alpha}5ß1 integrin by SNAKA51 increased the accumulation of detergent-resistant fibronectin in the extracellular matrix, thus identifying an integrin conformation that promotes matrix assembly. The SNAKA51 epitope was mapped to the calf-1/calf-2 domains. We propose that the action of the antibody causes the legs of the integrin to change conformation and thereby primes the integrin to bind ligand. These findings identify SNAKA51 as the first anti-integrin antibody to selectively recognize a subset of adhesion contacts, and they identify an integrin conformation associated with integrin translocation and fibronectin matrix formation.

Key words: Integrin, Fibronectin, Conformation, Monoclonal antibody, Matrix assembly




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