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Equilibrium and kinetics of the allosteric transition of GroEL Studied by solution X-ray scattering and fluorescence spectroscopy
https://ir.soken.ac.jp/records/4312
https://ir.soken.ac.jp/records/4312653062fe-6d7e-4974-ae5b-7362b4f9c840
| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||
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| 公開日 | 2014-03-19 | |||||||||||
| タイトル | ||||||||||||
| タイトル | Equilibrium and kinetics of the allosteric transition of GroEL Studied by solution X-ray scattering and fluorescence spectroscopy | |||||||||||
| タイトル | ||||||||||||
| タイトル | Equilibrium and kinetics of the allosteric transition of GroEL Studied by solution X-ray scattering and fluorescence spectroscopy | |||||||||||
| 言語 | en | |||||||||||
| 言語 | ||||||||||||
| 言語 | eng | |||||||||||
| 資源タイプ | ||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||
| 資源タイプ | journal article | |||||||||||
| アクセス権 | ||||||||||||
| アクセス権 | metadata only access | |||||||||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||||||||
| 著者 |
INOBE, Tomonao
× INOBE, Tomonao
× KUWAJIMA, Kunihiro
× et, al.
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| 著者別名 |
桑島, 邦博
× 桑島, 邦博
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| 抄録 | ||||||||||||
| 内容記述タイプ | Abstract | |||||||||||
| 内容記述 | We have studied the ATP-induced allosteric structural transition of GroEL using small angle X-ray scattering and fluorescence spectroscopy in combination with a stopped-flow technique. With X-ray scattering one can clearly distinguish the three allosteric states of GroEL, and the kinetics of the transition of GroEL induced by 85 μM ATP have been observed directly by stopped-flow X-ray scattering for the first time. The rate constant has been found to be 3–5 s−1 at 5 °C, indicating that this process corresponds to the second phase of the ATP-induced kinetics of tryptophan-inserted GroEL measured by stopped-flow fluorescence. Based on the ATP concentration dependence of the fluorescence kinetics, we conclude that the first phase represents bimolecular non-cooperative binding of ATP to GroEL with a bimolecular rate constant of 5.8×105 M−1 s−1 at 25 °C. Considering the electrostatic repulsion between negatively charged GroEL (−18 of the net charge per monomer at pH 7.5) and ATP, the rate constant is consistent with a diffusion-controlled bimolecular process. The ATP-induced fluorescence kinetics (the first and second phases) at various ATP concentrations (<400 μM) occur before ATP hydrolysis by GroEL takes place and are well explained by a kinetic allosteric model, which is a combination of the conventional transition state theory and the Monod–Wyman–Changeux model, and we have successfully evaluated the equilibrium and kinetic parameters of the allosteric transition, including the binding constant of ATP in the transition state of GroEL. | |||||||||||
| 書誌情報 |
Journal of Molecular Biology en : Journal of Molecular Biology 巻 327, 号 1, p. 183-191, 発行日 2003-03-14 |
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| 出版者 | ||||||||||||
| 出版者 | Elsevier | |||||||||||
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| 収録物識別子タイプ | ISSN | |||||||||||
| 収録物識別子 | 0022-2836 | |||||||||||
| DOI | ||||||||||||
| 識別子タイプ | DOI | |||||||||||
| 関連識別子 | https://doi.org/10.1016/S0022-2836(03)00087-1 | |||||||||||
| 関連名称 | 10.1016/S0022-2836(03)00087-1 | |||||||||||
| 権利 | ||||||||||||
| 権利情報 | © 2003 Elsevier Science Ltd. | |||||||||||
