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The tip then leaves contact with the surface and, as the ligand on the tip remains bound to surface receptor molecule, both are extended. With further application of retractionforce, the molecule-ligand complex dissociates (4). The cantilever then returns into its resting position, (I), and is ready for another measurement. The maximum diflerence between the approach and retraction curves, and the curve shape in the region of (4), gives information on the interaction ‘binding’force between the tip and surface and their physical properties.

These changes are triggered by alterations in the extracellular microenvironment of the receptor and by interactions between the cytoplasmic part of the receptor and intracellular signalling molecules and the cytoskeleton. It is likely that this is true for many other systems of cell surface membrane receptors. RGD binding forces, measured by AFM in live bone cells, osteoclasts and osteoblasts, is dependent not only on the type of integrin expressed by a cell and its activation status but also on the spatial conformation of the sequence in which the RGD sequence is present54This may be because a more favorable orientation between the ligand and the receptor results in shorter interaction ranges for the molecules and thus higher interaction forces between RGD and the binding site.

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