HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 283, Issue 35, 23765-23773, August 29, 2008

This Article Full Text Full Text (PDF) Author Profile Supplemental Data All Versions of this Article: 283/35/23765    most recent M802541200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sameshima, T. Articles by Funatsu, T. Search for Related Content PubMed PubMed Citation Articles by Sameshima, T. Articles by Funatsu, T. Related Collections Papers Of The Week Social Bookmarking                      What's this?

Football- and Bullet-shaped GroEL-GroES Complexes Coexist during the Reaction Cycle^*

Tomoya Sameshima, Taro Ueno, Ryo Iizuka¹, Noriyuki Ishii², Naofumi Terada³, Kohki Okabe, and Takashi Funatsu^¶4

From the Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan, the Biological Information Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba Central-6, 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8566, Japan, and the ^¶Center for NanoBio Integration, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan

GroEL is an Escherichia coli chaperonin that is composed of two heptameric rings stacked back-to-back. GroEL assists protein folding with its cochaperonin GroES in an ATP-dependent manner in vitro and in vivo. However, it is still unclear whether GroES binds to both rings of GroEL simultaneously under physiological conditions. In this study, we monitored the GroEL-GroES interaction in the reaction cycle using fluorescence resonance energy transfer. We found that nearly equivalent amounts of symmetric GroEL-(GroES)₂ (football-shaped) complex and asymmetric GroEL-GroES (bullet-shaped) complex coexist during the functional reaction cycle. We also found that D398A, an ATP hydrolysis defective mutant of GroEL, forms a football-shaped complex with ATP bound to the two rings. Furthermore, we showed that ADP prevents the association of ATP to the trans-ring of GroEL, and as a consequence, the second GroES cannot bind to GroEL. Considering the concentrations of ADP and ATP in E. coli, ADP is expected to have a small effect on the inhibition of GroES binding to the trans-ring of GroEL in vivo. These results suggest that we should reconsider the chaperonin-mediated protein-folding mechanism that involves the football-shaped complex.

Received for publication, April 2, 2008 , and in revised form, June 12, 2008.

^* This work was supported by a grant from the Mitsubishi Foundation (to T. F.) and by Grant-in-Aid for Scientific Research (A) 17201031 (to T. F.) from the Ministry of Education, Culture, Sports, Science and Technology 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental text and five supplemental figures.

This article was selected as a Paper of the Week.

¹ The recipient of a Research Fellowship from the Japan Society for the Promotion of Science for Young Scientists (18·10141).

² Present address: Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central-6, 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8566, Japan.

³ Present address: RIKEN (The Institute of Physical and Chemical Research), 2-1, Hirosawa, Wako-shi, Saitama 351-0198, Japan.

⁴ 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.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. Koike-Takeshita, M. Yoshida, and H. Taguchi Revisiting the GroEL-GroES Reaction Cycle via the Symmetric Intermediate Implied by Novel Aspects of the GroEL(D398A) Mutant J. Biol. Chem., August 29, 2008; 283(35): 23774 - 23781. [Abstract] [Full Text] [PDF]