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*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)


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
Issue number30
Publication statusPublished - 2010 Jul 23

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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