| Item type |
学術雑誌論文 / Journal Article(1) |
| 公開日 |
2014-03-20 |
| タイトル |
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タイトル |
Acid Denaturation and Refolding of Green Fluorescent Protein |
| タイトル |
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タイトル |
Acid Denaturation and Refolding of Green Fluorescent Protein |
|
言語 |
en |
| 言語 |
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|
言語 |
eng |
| 資源タイプ |
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資源タイプ識別子 |
http://purl.org/coar/resource_type/c_6501 |
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資源タイプ |
journal article |
| アクセス権 |
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アクセス権 |
metadata only access |
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アクセス権URI |
http://purl.org/coar/access_right/c_14cb |
| 著者 |
ENOKI, Sawako
SAEKI, Kimiko
MäKI-PAAKKANEN, Jorma
KUWAJIMA, Kunihiro
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| 著者別名 |
桑島, 邦博
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| 抄録 |
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内容記述タイプ |
Abstract |
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内容記述 |
Green fluorescent protein from the jellyfish Aequorea victoria can serve as a good model protein to understand protein folding in a complex environment with molecular chaperones and other macromolecules such as those in biological cells, but little is known about the detailed mechanisms of the in vitro folding of green fluorescent protein itself. We therefore investigated the kinetic refolding of a mutant (F99S/M153T/V163A) of green fluorescent protein, which is known to mature more efficiently than the wild-type protein, from the acid-denatured state; refolding was observed by chromophore fluorescence, tryptophan fluorescence, and far-UV CD, using a stopped-flow technique. In this study, we demonstrated that the kinetics of the refolding of the mutant have at least five kinetic phases and involve nonspecific collapse within the dead time of a stopped-flow apparatus and the subsequent formation of an on-pathway intermediate with the characteristics of the molten globule state. We also demonstrated that the slowest phase and a major portion of the second slowest phase were rate-limited by slow prolyl isomerization in the intermediate state, and this rate limitation accounts for a major portion of the observed kinetics in the folding of green fluorescent protein. |
| 書誌情報 |
Biochemistry
en : Biochemistry
巻 43,
号 44,
p. 14238-14248,
発行日 2004
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| 出版者 |
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出版者 |
American Chemical Society |
| ISSN |
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収録物識別子タイプ |
ISSN |
|
収録物識別子 |
0006-2960 |
| DOI |
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識別子タイプ |
DOI |
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関連識別子 |
http://doi.org/10.1021/bi048733+ |
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関連名称 |
10.1021/bi048733+ |
| 権利 |
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権利情報 |
© 2004 American Chemical Society |
