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The Burst-phase Intermediate in the Refolding of β-Lactoglobulin Studied by Stopped-flow Circular Dichroism and Absorption Spectroscopy
https://ir.soken.ac.jp/records/4270
https://ir.soken.ac.jp/records/427063dd68f9-53cd-4550-9c03-266cb91dd494
| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||
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| 公開日 | 2014-03-10 | |||||||||||
| タイトル | ||||||||||||
| タイトル | The Burst-phase Intermediate in the Refolding of β-Lactoglobulin Studied by Stopped-flow Circular Dichroism and Absorption Spectroscopy | |||||||||||
| タイトル | ||||||||||||
| タイトル | The Burst-phase Intermediate in the Refolding of β-Lactoglobulin Studied by Stopped-flow Circular Dichroism and Absorption 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 |
KUWAJIMA, Kunihiro
× KUWAJIMA, Kunihiro
× YAMAYA, Hidetoshi
× SUGAI, Shintaro
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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 | The kinetics of the guanidine hydrochloride-induced unfolding and refolding of bovine β-lactoglobulin, a predominantly β-sheet protein in the native state, have been studied by stopped-flow circular dichroism and absorption measurements at pH 3.2 and 4.5°C. The refolding reaction was a complex process composed of different kinetic phases, while the unfolding was a single-phase reaction. Most notably, a burst-phase intermediate of refolding, which was formed during the dead time of stopped-flow measurements (∼18 ms), showed more intense ellipticity signals in the peptide region below 240 nm than the native state, yielding overshoot behavior in the refolding curves. We have investigated the spectral properties and structural stability of the burst-phase intermediate and also the structural properties in the unfolded state in 4.0 M guanidine hydrochloride of the protein and its disulfide-cleaved derivative. The main conclusions are: (1) the more intense ellipticity of the intermediate in the peptide region arises from formation of non-native α-helical structure in the intermediate, apparently suggesting that the folding of β-lactoglobulin is not represented by a simple sequential mechanism. (2) The burst-phase intermediate has, however, a number of properties in common with the folding intermediates or with the molten globule states of other globular proteins whose folding reactions are known to be represented by the sequential model. These properties include: the presence of the secondary structure without the specific tertiary structure; formation of a hydrophobic core; broad unfolding transition of the intermediate; and rapidity of formation of the intermediate. The burst-phase intermediate of β-lactoglobulin is thus classified as the same species as the molten globule state. (3) The circular dichroism spectra of β-lactoglobulin and its disulfide-cleaved derivative in 4.0 M guanidine hydrochloride suggests the presence of the residual β-structure in the unfolded state and the stabilization of the β-structure by disulfide bonds. Thus, if this residual β-structure is part of the native β-structure and forms a folding initiation site, the folding reaction of β-lactoglobulin may not necessarily be inconsistent with the sequential model. The non-native α-helices in the burst-phase intermediate may be formed in an immature part of the protein molecule because of the local α-helical propensity in this part. | |||||||||||
| 書誌情報 |
Journal of Molecular Biology en : Journal of Molecular Biology 巻 264, 号 4, p. 806-822, 発行日 1996-12-13 |
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| 出版者 | ||||||||||||
| 出版者 | Elsevier | |||||||||||
| ISSN | ||||||||||||
| 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/10.1006/jmbi.1996.0678 | |||||||||||
| 関連名称 | 10.1006/jmbi.1996.0678 | |||||||||||
| 権利 | ||||||||||||
| 権利情報 | © 1996 Academic Press | |||||||||||
