Single-molecule study on the decay process of the football-shaped GroEL-GroES complex using zero-mode waveguides

Tomoya Sameshima, Ryo Iizuka, Taro Ueno, Junichi Wada, Mutsuko Aoki, Naonobu Shimamoto, Iwao Ohdomari, Takashi Tanii, Takashi Funatsu

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    It has been widely believed that an asymmetric GroEL-GroES complex (termed the bullet-shaped complex) is formed solely throughout the chaperonin reaction cycle, whereas we have recently revealed that a symmetric GroEL-(GroES) 2 complex (the football-shaped complex) can form in the presence of denatured proteins. However, the dynamics of the GroEL-GroES interaction, including the football-shaped complex, is unclear. We investigated the decay process of the football-shaped complex at a single-molecule level. Because submicromolar concentrations of fluorescent GroES are required in solution to form saturated amounts of the football-shaped complex, single-molecule fluorescence imaging was carried out using zero-mode waveguides. The single-molecule study revealed two insights into the GroEL-GroES reaction. First, the first GroES to interact with GroEL does not always dissociate from the football-shaped complex prior to the dissociation of a second GroES. Second, there are two cycles, the "football cycle" and the "bullet cycle," in the chaperonin reaction, and the lifetimes of the football-shaped and the bullet-shaped complexes were determined to be 3-5 s and about 6 s, respectively. These findings shed new light on the molecular mechanism of protein folding mediated by the GroEL-GroES chaperonin system.

    Original languageEnglish
    Pages (from-to)23159-23164
    Number of pages6
    JournalJournal of Biological Chemistry
    Volume285
    Issue number30
    DOIs
    Publication statusPublished - 2010 Jul 23

    Fingerprint

    Chaperonins
    Football
    Waveguides
    Molecules
    Protein folding
    Fluorescence
    Imaging techniques
    Optical Imaging
    Protein Folding
    Proteins

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Cite this

    Sameshima, T., Iizuka, R., Ueno, T., Wada, J., Aoki, M., Shimamoto, N., ... Funatsu, T. (2010). Single-molecule study on the decay process of the football-shaped GroEL-GroES complex using zero-mode waveguides. Journal of Biological Chemistry, 285(30), 23159-23164. https://doi.org/10.1074/jbc.M110.122101

    Single-molecule study on the decay process of the football-shaped GroEL-GroES complex using zero-mode waveguides. / Sameshima, Tomoya; Iizuka, Ryo; Ueno, Taro; Wada, Junichi; Aoki, Mutsuko; Shimamoto, Naonobu; Ohdomari, Iwao; Tanii, Takashi; Funatsu, Takashi.

    In: Journal of Biological Chemistry, Vol. 285, No. 30, 23.07.2010, p. 23159-23164.

    Research output: Contribution to journalArticle

    Sameshima, T, Iizuka, R, Ueno, T, Wada, J, Aoki, M, Shimamoto, N, Ohdomari, I, Tanii, T & Funatsu, T 2010, 'Single-molecule study on the decay process of the football-shaped GroEL-GroES complex using zero-mode waveguides', Journal of Biological Chemistry, vol. 285, no. 30, pp. 23159-23164. https://doi.org/10.1074/jbc.M110.122101
    Sameshima, Tomoya ; Iizuka, Ryo ; Ueno, Taro ; Wada, Junichi ; Aoki, Mutsuko ; Shimamoto, Naonobu ; Ohdomari, Iwao ; Tanii, Takashi ; Funatsu, Takashi. / Single-molecule study on the decay process of the football-shaped GroEL-GroES complex using zero-mode waveguides. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 30. pp. 23159-23164.
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