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Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy

Nature volume 397, pages 50–53 (1999)Cite this article

Abstract

Atomic force microscopy (AFM)1, 2 has been used to measure the strength of bonds between biological receptor molecules and their ligands3,4,5,6. But for weak noncovalent bonds, a dynamic spectrum of bond strengths is predicted as the loading rate is altered, with the measured strength being governed by the prominent barriers traversed in the energy landscape along the force-driven bond-dissociation pathway7. In other words, the pioneering early AFM measurements represent only a single point in a continuous spectrum of bond strengths, because theory predicts that these will depend on the rate at which the load is applied. Here we report the strength spectra for the bonds between streptavidin (oravidin) and biotin8—the prototype of receptor–ligand interactions used in earlier AFM studies3,4,5, and which have been modelled by molecular dynamics9, 10. We have probed bond formation over six orders of magnitude in loading rate, and find that the bond survival time diminished from about 1 min to 0.001 s with increasing loading rate over this range. The bond strength, meanwhile, increased from about 5 pN to 170 pN. Thus, although they are among the strongest noncovalent linkages in biology (affinity of 1013 to 1015 M−1)8, 11, these bonds in fact appear strong or weak depending on how fast they are loaded. We are also able to relate the activation barriers derived from our strength spectra to the shape of the energy landscape derived from simulations of the biotin–avidin complex.

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Figure 1: The spring in the biomembrane force probe BFP is a pressurized membrane capsule12.
Figure 2: BFP tip–substrate distance and force versus time for cycles of approach–touch–separation with formation and rupture of a bond.
Figure 3: Biotin–streptavidin bond strengths.
Figure 4: Conceptual and real (MD) energy landscapes traversed along a molecular reaction coordinate under force.

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Acknowledgements

We thank A. Chilkoti, C. Cantor and the group of K. Schulten for helpful discussions. The work was supported by USPHS National Institutes of Health, Medical Research Council of Canada, and the Canadian Institute for Advanced Research Program in Science of Soft Surfaces and Interfaces.

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Authors and Affiliations

  1. Departments of Physics and Pathology, University of British Columbia, Vancouver, V6T 1Z1, British Columbia , Canada

    R. Merkel, P. Nassoy, A. Leung, K. Ritchie & E. Evans

  2. Physikdepartment der Technischen Universitt Mnchen, 85748 Garching, Germany

    R. Merkel

  3. Physico-Chemie de l'Institut Curie , 75231, Paris, France

    P. Nassoy

  4. Biomedical Engineering, Boston University , Boston, 02215, Massachusetts, USA

    E. Evans

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Correspondence to E. Evans.

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Merkel, R., Nassoy, P., Leung, A. et al. Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy. Nature 397, 50–53 (1999). https://doi.org/10.1038/16219

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