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Probing protein–protein interactions in real time

Nature Structural Biology volume 7, pages 644–647 (2000)Cite this article

Abstract

We have used a prototype small cantilever atomic force microscope to observe, in real time, the interactions between individual protein molecules. In particular, we have observed individual molecules of the chaperonin protein GroES binding to and then dissociating from individual GroEL proteins, which were immobilized on a mica support. This work suggests that the small cantilever atomic force microscope is a useful tool for studying protein dynamics at the single molecule level.

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Figure 1: Small cantilevers and image of GroEL taken with the small cantilever AFM.
Figure 2: GroEL and isolated GroEL–GroES complexes in buffer solution on a mica support.
Figure 3: GroEL scanned in two dimensions (left) and in one dimension (right).
Figure 4: Association and dissociation of the GroEL–GroES complex.
Figure 5: Measured complex lifetime of an individual GroEL molecule in the presence of Mg-ATP and GroES.

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Acknowledgements

We thank G. Lorimer for his encouragement and many useful suggestions. We thank H. Saibil for her generous permission to use the cryo-electron microscopy images of GroEL and the GroEL–GroES complex. The Materials Research Division and the Molecular Biophysics Division of the National Science Foundation supported this work.

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Authors and Affiliations

  1. Department of Physics, University of California at Santa Barbara, Santa Barbara, 93106, California, USA

    Mario B. Viani, Lia I. Pietrasanta, James B. Thompson, Ami Chand, Ilse C. Gebeshuber, Johannes H. Kindt, Michael Richter, Helen G. Hansma & Paul K. Hansma

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  1. Mario B. Viani
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Correspondence to Mario B. Viani.

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Viani, M., Pietrasanta, L., Thompson, J. et al. Probing protein–protein interactions in real time. Nat Struct Mol Biol 7, 644–647 (2000). https://doi.org/10.1038/77936

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