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Single-molecule observation of protein–protein interactions in the chaperonin system

Nature Biotechnology volume 19, pages 861–865 (2001)Cite this article

Abstract

We have analyzed the dynamics of the chaperonin (GroEL)–cochaperonin (GroES) interaction at the single-molecule level. In the presence of ATP and non-native protein, binding of GroES to the immobilized GroEL occurred at a rate that is consistent with bulk kinetics measurements. However, the release of GroES from GroEL occurred after a lag period (∼3 s) that was not recognized in earlier bulk-phase studies. This observation suggests a new kinetic intermediate in the GroEL–GroES reaction pathway.

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Figure 1: Single-molecule imaging of chaperonin GroEL–GroES dynamics.
Figure 2: Statistical analysis of GroES association and dissociation with GroEL in the presence of reduced lactalbumin.
Figure 3: (A) Effect of ATP concentration on dissociation of GroES.

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Acknowledgements

This work was partly supported by the Yamada Foundation (H.T. and T.F.) and a grant-in-aid for Scientific Research on Priority Areas (A) from the Ministry of Education, Science, Sports and Culture of Japan (H.T., M.Y., and T.F.). We thank Dr. A. Horwich for discussions, Dr. F. Motojima for discussion of polypeptide–GroEL binding, and Dr. J. Hardy for critical reading of the manuscript. We also thank Dr. K. Aoki for providing polypeptide RP3-42 and Mrs. J. Suzuki for technical assistance.

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Author notes

  1. Hideki Taguchi, Taro Ueno and Hisashi Tadakuma: These three authors contributed equally to this work.

Authors and Affiliations

  1. Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsuta, Yokohama, 226-8503, Japan

    Hideki Taguchi & Masasuke Yoshida

  2. Department of Physics, School of Science and Engineering, Waseda University, 3-4-1 Okubo, Tokyo, 169-8555, Japan

    Taro Ueno, Hisashi Tadakuma & Takashi Funatsu

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  1. Hideki Taguchi
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  2. Taro Ueno
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  4. Masasuke Yoshida
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  5. Takashi Funatsu
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Corresponding author

Correspondence to Masasuke Yoshida.

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Taguchi, H., Ueno, T., Tadakuma, H. et al. Single-molecule observation of protein–protein interactions in the chaperonin system. Nat Biotechnol 19, 861–865 (2001). https://doi.org/10.1038/nbt0901-861

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