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Direct Observation of Multiple and Stochastic Transition States by a Feedback-controlled Single-molecule Force Measurement

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Abstract

To overcome the ensemble-averaging barrier, single-molecule experiments have been performed, but energy landscapes comprising multiple intermediates have not yet been defined. We performed mechanical unfolding of staphylococcal nuclease using intermolecular force microscopy, modified AFM with high resolution and feedback control of the positioning. The force dropped vertically just after its peak, and multiple transition states were detected as force peaks. The multiple and stochastic intermediates found in the present study provide new important information on protein folding.

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Authors and Affiliations

  1. Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka, 411-8540, Japan

    Akihiro Fukagawa, Michio Hiroshima, Isao Sakane & Makio Tokunaga

  2. Research Center for Allergy and Immunology, RIKEN, Yokohama, Kanagawa, 230-0045, Japan

    Akihiro Fukagawa, Michio Hiroshima & Makio Tokunaga

  3. Department of Genetics, School of Life Science, The Graduate University for Advanced Studies (Sokendai), Mishima, Shizuoka, 411-8540, Japan

    Makio Tokunaga

  4. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori, Yokohama, Kanagawa, 226-8501, Japan

    Makio Tokunaga

Authors
  1. Akihiro Fukagawa
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  2. Michio Hiroshima
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  3. Isao Sakane
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  4. Makio Tokunaga
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Correspondence to Makio Tokunaga.

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Fukagawa, A., Hiroshima, M., Sakane, I. et al. Direct Observation of Multiple and Stochastic Transition States by a Feedback-controlled Single-molecule Force Measurement. ANAL. SCI. 25, 5–7 (2009). https://doi.org/10.2116/analsci.25.5

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  • DOI: https://doi.org/10.2116/analsci.25.5

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