title: Single integrin molecule adhesion forces in intact cells measured by atomic force microscopy.
abstract: Cross-talk between cells and the extracellular matrix is critically influenced by the mechanical properties of cell surface receptor-ligand interactions; these interactions are especially well defined and regulated in cells capable of dynamically modifying their matrix environment. In this study, attention was focused on osteoclasts, which are absolutely dependent on integrin extracellular matrix receptors in order to degrade bone; other bone cells, osteoblasts, were used for comparison. Integrin binding forces were measured in intact cells by atomic force microscopy (AFM) for several RGD-containing (Arg-Gly-Asp) ligands and ranged from 32 to 97 picoNewtons (pN); they were found to be cell and amino acid sequence specific, saturatable and sensitive to the pH and divalent cation composition of the cellular culture medium. In contrast to short linear RGD hexapeptides, larger peptides and proteins containing the RGD sequence, such as osteopontin (a major non-collagenous bone protein) and echistatin (a high affinity RGD sequence containing antagonist snake venom protein), showed different binding affinities. This demonstrates that the context of the RGD sequence within a protein has considerable influence upon the final binding force for receptor interaction. These data also demonstrate that AFM, as a methodological approach, can be adapted to cell biology studies wherever cell-matrix interactions play a critical role, and, moreover, may have applicability to the analysis of receptor-ligand interactions in cell membranes in general.
authors.affiliation: Department of Medicine, The Rayne Institute, 5 University Street, University College London, WC1E 6JJ, United Kingdom.
authors.forename: P P
authors.initials: PP
authors.lastname: Lehenkari
authors.forename: M A
authors.initials: MA
authors.lastname: Horton
mesh_terms: D000818:Animals; D016023:Integrins / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D010010:Osteoclasts / Q000737:chemistry*; D011485:Protein Binding; D051381:Rats
chemical_list: D016023:Integrins