Abstract
Cell–cell adhesion mediated by specific cell-surface molecules is essential for multicellular development. Here we quantify de-adhesion forces at the resolution of individual cell-adhesion molecules, by controlling the interactions between single cells and combining single-molecule force spectroscopy with genetic manipulation. Our measurements are focused on a glycoprotein, contact site A (csA), as a prototype of cell-adhesion proteins. csA is expressed in aggregating cells of Dictyostelium discoideum, which are engaged in development of a multicellular organism. Adhesion between two adjacent cell surfaces involves discrete interactions characterized by an unbinding force of 23 ± 8 pN, measured at a rupture rate of 2.5 ± 0.5 µm s–1.
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References
Vestweber, D. & Blanks, J. E. Mechanisms that regulate the function of the selectins and their ligands. Physiol. Rev. 79 , 181–213 (1999).
Fritz, J., Katopodis, A. G., Kolbinger, F. & Anselmetti, D. Force-mediated kinetics of single P-selectin/ligand complexes observed by atomic force microscopy. Proc. Natl Acad. Sci. USA 95, 12283–12288 (1998).
Springer, T. A. Adhesion receptors of the immune system. Nature 346 , 425–434 (1990).
Kreis, T. & Vale, R. (eds) Guidebook to the Extracellular Matrix, Anchor, and Adhesion Proteins (Oxford Univ. Press, 1999).
Faix, J. in Guidebook to the Extracellular Matrix, Anchor, and Adhesion Proteins (eds Kreis, T. & Vale, R.) 177–179 (Oxford Univ. Press, 1999).
Ponte, E., Bracco, E., Faix, J. & Bozzaro, S. Detection of subtle phenotypes: the case of the cell adhesion molecule csA in Dictyostelium . Proc. Natl Acad. Sci. USA 95, 9360– 9365 (1998).
Bozzaro, S. & Gerisch, G. Contact sites in aggregating cells of Polysphondylium pallidum. J. Mol. Biol. 120, 265–279 (1978).
Gerisch, G., Krelle, H., Bozzarro, S., Eitle, E. & Guggenheim, R. in Cell Adhesion and Motility (eds Curtis, A. S. G. & Pitts, J. D.) 293–307 (Cambridge Univ. Press, Cambridge, UK, 1980).
Murray, B. A., Yee, L. D. & Loomis, W. F. Immunological analysis of glycoprotein (contact sites A) involved in intercellular adhesion of Dictyostelium discoideum. J. Supramol. Struct. Cell. Biochem. 17, 197– 211 (1981).
Beug, H., Katz, F. E. & Gerisch, G. Dynamics of antigenic membrane sites relating to cell aggregation in Dictyostelium discoideum. J. Cell Biol. 56, 647–658 (1973).
Siu, C.H. et al. Molecular mechanisms of cell–cell interaction in Dictyostelium discoideum. Biochem. Cell Biol. 66, 1089–1099 (1988).
Stadler, J., Keenan, T. W., Bauer, G. & Gerisch, G. The contact site A glycoprotein of Dictyostelium discoideum carries a phospholipid anchor of a novel type. EMBO J. 8, 371– 377 (1989).
Barth, A., Müller-Taubenberger, A., Taranto, P. & Gerisch, G. Replacement of the phospholipid-anchor in the contact site A glycoprotein of Dictyostelium discoideum by a transmembrane region does not impede cell adhesion but reduces residence time on the cell surface. J. Cell Biol. 124, 205– 215 (1994).
Evans, E. A. Detailed mechanics of membrane–membrane adhesion and separation II. Discrete kinetically trapped molecular cross-bridges. Biophys. J. 48, 185–192 ( 1985).
Evans, E. in Structure and Dynamics of Membranes (eds Lipowsky, R. & Sackmann, E.) 723–754 (Elsevier, Amsterdam, 1995).
Curtis, A. S. G. Problems and some solutions in the study of cellular aggregation. Symp. Zool. Soc. Lond. 25, 335–352 (1970).
Chen, S. & Springer, T. A. An automatic braking system that stabilizes leukocyte rolling by an increase in selectin bond number with shear. J. Cell Biol. 144, 185– 200 (1999).
Binnig, G., Quate, C. F. & Gerber, C. Atomic force microscope. Phys. Rev. Lett. 56, 930–933 ( 1986).
Gimzewski, J. K. & Joachim, C. Nanoscale science of single molecules using local probes. Science 283 , 1683–1688 (1999).
Oesterhelt, F. et al. Unfolding pathways of individual bacteriorhodopsins. Science (in the press).
Rief, M., Oesterhelt, F., Heymann, B. & Gaub, H. E. Single molecule force spectroscopy on polysaccharides by AFM. Science 275, 1295–1297 ( 1997).
Marszalek, P. E. et al. Atomic levers control pyranose ring conformations. Proc. Natl Acad. Sci. USA 96, 7894– 7898 (1999).
Rief, M., Gautel, M., Schemmel, A. & Gaub, H. E. The mechanical stability of immunoglobulin and fibronectin III domains in the muscle protein titin measured by AFM. Biophys. J. 75, 3008 –3014 (1998).
Oberhauser, A. F., Marszalek, P. E., Erickson, H. P. & Fernandez, J. M. The molecular elasticity of the extracellular matrix protein tenascin. Nature 393, 181–185 ( 1998).
Smith, B. L. et al. Molecular mechanistic origin of the toughness of natural adhesives, fibres and composites. Nature 399, 761– 763 (1999).
Rief, M., Clausen-Schaumann, H. & Gaub, H. E. Sequence-dependent mechanics of single DNA molecules . Nature Struct. Biol. 6, 346– 349 (1999).
Strunz, T., Oroszlan, K., Schäfer, R. & Güntherodt, H-J. Dynamic force spectroscopy of single DNA molecules. Proc. Natl Acad. Sci. USA 96, 11277–11282 (1999).
Moy, V. T., Florin, E-L. & Gaub, H. E. Intermolecular forces and energies between ligands and receptors. Science 266, 257–259 (1994).
Florin, E-L., Moy, V. T. & Gaub, H. E. Adhesive forces between individual ligand-receptor pairs . Science 264, 415–417 (1994).
Hinterdorfer, P., Baumgartner, W., Gruber, H. J., Schilcher, K. & Schindler, H. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy . Proc. Natl Acad. Sci. USA 93, 3477– 3481 (1996).
Müller, K. M., Arndt, K. M. & Plückthun, A. Model and simulation of multivalent binding to fixed ligands. Anal. Biochem. 261, 149–158 ( 1998).
Grubmüller, H., Heymann, B. & Tavan, P. Ligand binding: molecular mechanics calculation of the streptavidin–biotin rupture force. Science 271 , 997–999 (1996).
Merkel, R., Nassoy, P., Leung, A., Ritchie, K. & Evans, E. Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy. Nature 397, 50–53 (1999).
Beug, H., Katz, F. E., Stein, A. & Gerisch, G. Quantitation of membrane sites in aggregating Dictyostelium cells by use of tritiated univalent antibody. Proc. Natl Acad. Sci. USA 70, 3150– 3154 (1973).
Harloff, C., Gerisch, G. & Noegel, A. A. Selective elimination of the contact site A protein of Dictyostelium discoideum by gene disruption. Genes Dev. 3, 2011–2019 (1989).
Faix, J., Gerisch, G. & Noegel, A. A. Overexpression of the csA cell adhesion molecule under its own cAMP-regulated promoter impairs morphogenesis in Dictyostelium . J. Cell Sci. 102, 203–214 (1992).
Faix, J., Gerisch, G. & Noegel, A. A. Constitutive overexpression of the contact site A glycoprotein enables growth-phase cells of Dictyostelium discoideum to aggregate. EMBO J. 9, 2709–2716 ( 1990).
Evans, E. Energy landscapes of biomolecular adhesion and receptor anchoring at interfaces explored with dynamic force spectroscopy. R. Soc. Chem: Faraday Discuss. 111, 1–16 ( 1998).
Tomschy, A., Fauser, C., Landwehr, R. & Engel, J. Homophilic adhesion of E-cadherin occurs by a co-operative two-step interaction of N-terminal domains. EMBO J. 15, 3507– 3514 (1996).
Rief, M., Fernandez, J. M. & Gaub, H. E. Elastically coupled two-level systems as a model for biopolymer extensibility. Phys. Rev. Lett. 81, 4764–4767 (1998).
Evans, E., Berk, D. & Leung, A. Detachment of agglutinin bonded red blood cells I. Forces to rupture molecular point attachments. Biophys. J. 59, 838– 848 (1991).
Bell, G. I. Models for the specific adhesion of cells to cells. Science 200, 618–627 (1978).
Dettmann, W. et al. Differences in zero-force and force-driven kinetics of ligand dissociation from β-galactoside-specific proteins (plant and animal lectins, immunoglobulin G) monitored by plasmon resonance and dynamic single molecule force microscopy. Arch. Biochem. Biophys. (in the press).
Kloboucek, A., Behrisch, A., Faix, J. & Sackmann, E. Adhesion-induced receptor segregation and adhesion plaque formation: a model membrane study . Biophys. J. 77, 2311– 2328 (1999).
Malchow, D., Nägele, B., Schwarz, H. & Gerisch, G. Membrane-bound cyclic AMP phosphodiesterase in chemotactically responding cells of Dictyostelium discoideum. Eur. J. Biochem. 28, 136–142 (1972).
Florin, E. L. et al. Sensing specific molecular interactions with the atomic force microscope. Biosensors Bioelectr. 10, 895 –901 (1995).
Butt, H. J. & Jaschke, M. Thermal noise in atomic force microscopy . Nanotechnol. 6, 1–7 (1995).
Acknowledgements
We thank M. Westphal and E. Simmeth for culturing Dictyostelium, and W. Dettmann for instrumentation and discussions. This work was supported by grants from the VW Foundation and the Deutsche Forschungsgemeinschaft..
Correspondence and requests for materials should be addressed to M.B.
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Benoit, M., Gabriel, D., Gerisch, G. et al. Discrete interactions in cell adhesion measured by single-molecule force spectroscopy. Nat Cell Biol 2, 313–317 (2000). https://doi.org/10.1038/35014000
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DOI: https://doi.org/10.1038/35014000
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