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J. Biol. Chem., Vol. 283, Issue 35, 23931-23939, August 29, 2008

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Effect of the C-terminal Truncation on the Functional Cycle of Chaperonin GroEL

IMPLICATION THAT THE C-TERMINAL REGION FACILITATES THE TRANSITION FROM THE FOLDING-ARRESTED TO THE FOLDING-COMPETENT STATE^*

Mihoko Suzuki, Taro Ueno, Ryo Iizuka¹, Takahiro Miura, Tamotsu Zako^¶, Rena Akahori, Takeo Miyake, Naonobu Shimamoto^||, Mutsuko Aoki, Takashi Tanii^||, Iwao Ohdomari^||, and Takashi Funatsu^**2

From the Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, the Faculty of Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, the ^¶Bioengineering Laboratory, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, the ^||Nanotechnology Research Center, Waseda University, 513 Tsurumaki-chou, Shinjuku-ku, Tokyo 162-0041, and the ^**Center for NanoBio Integration, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan

To elucidate the exact role of the C-terminal region of GroEL in its functional cycle, the C-terminal 20-amino acid truncated mutant of GroEL was constructed. The steady-state ATPase rate and duration of GroES binding showed that the functional cycle of the truncated GroEL is extended by 2 s in comparison with that of the wild type, without interfering with the basic functions of GroEL. We have proposed a model for the functional cycle of GroEL, which consists of two rate-limiting steps of 3- and 5-s duration ( Ueno, T., Taguchi, H., Tadakuma, H., Yoshida, M., and Funatsu, T. (2004) Mol. Cell 14, 423-434[CrossRef][Medline] [Order article via Infotrieve] g). According to the model, detailed kinetic studies were performed. We found that a 20-residue truncation of the C terminus extends the time until inorganic phosphate is generated and the time for arresting protein folding in the central cavity, i.e. the lifetime of the first rate-limiting step in the functional cycle, to an 5-s duration. These results suggest that the integrity of the C-terminal region facilitates the transition from the first to the second rate-limiting state.

Received for publication, May 29, 2008 , and in revised form, June 25, 2008.

^* This work was supported in part by Takeda Science Foundation (to T. F.), Grant-in-aid for Scientific Research (A) 17201031 (to T. F.), Grant-in-aid for Young Scientists 18770126 (to T. U.), and Grant-in-aid for Scientific Research on Priority Areas 18031008 (to T. U.) from the Ministry of Education, Science, Sports and Culture of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Recipient of Research Fellowship 18-10141 from the Japan Society for the Promotion of Science for Young Scientists.

² To whom correspondence should be addressed. Tel.: 81-3-5841-4760; Fax: 81-3-5802-3339; E-mail: funatsu{at}mail.ecc.u-tokyo.ac.jp.

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