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Folding of Green Fluorescent Protein and the Cycle3 Mutant
https://ir.soken.ac.jp/records/4296
https://ir.soken.ac.jp/records/429689755c3d-65e6-4f94-86c4-44582b29a33b
| Item type | 学術雑誌論文 / Journal Article(1) | |||||||||||
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| 公開日 | 2014-03-14 | |||||||||||
| タイトル | ||||||||||||
| タイトル | Folding of Green Fluorescent Protein and the Cycle3 Mutant | |||||||||||
| タイトル | ||||||||||||
| タイトル | Folding of Green Fluorescent Protein and the Cycle3 Mutant | |||||||||||
| 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 |
FUKUDA, Hiroyuki
× FUKUDA, Hiroyuki
× ARAI, Munehito
× KUWAJIMA, Kunihiro
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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 | Although the correct folding of green fluorescent protein (GFP) is required for formation of the chromophore, it is known that wild-type GFP cannot mature efficiently in vivo in Escherichia coli at 37 °C or higher temperatures that the jellyfish in the Pacific Northwest have never experienced. Recently, by random mutagenesis by the polymerase chain reaction (PCR) method, a mutant called Cycle3 was constructed. This mutant had three mutations, F99S, M153T, and V163A, on or near the surface of the GFP molecule and was able to mature correctly even at 37 °C [Crameri et al. (1996) Nat. Biotechnol. 143, 315−319]. In the present study, we investigated the differences in their folding behavior in vitro. We observed the folding and unfolding reactions of both wild-type GFP and the Cycle3 mutant by using green fluorescence as an indicator of the formation of the native structure and examining hydrogen-exchange reactions by Fourier transform infrared spectroscopy. Both proteins showed unusually slow refolding and unfolding rates, and their refolding rates were almost identical under the native state at 25 and at 35 °C. On the other hand, aggregation studies in vitro showed that wild-type GFP had a strong tendency to aggregate, while the Cycle3 mutant did not. These results indicated that the ability to mature efficiently in vivo at 37 °C was not due to the improved folding and that reduced hydrophobicity on the surface of the Cycle3 mutant was a more critical factor for efficient maturation in vivo. | |||||||||||
| 書誌情報 |
Biochemistry en : Biochemistry 巻 39, 号 39, p. 12025-12032, 発行日 2000 |
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| 出版者 | ||||||||||||
| 出版者 | American Chemical Society | |||||||||||
| ISSN | ||||||||||||
| 収録物識別子タイプ | ISSN | |||||||||||
| 収録物識別子 | 0006-2960 | |||||||||||
| DOI | ||||||||||||
| 識別子タイプ | DOI | |||||||||||
| 関連識別子 | http://doi.org/10.1021/bi000543l | |||||||||||
| 関連名称 | 10.1021/bi000543l | |||||||||||
| 権利 | ||||||||||||
| 権利情報 | © 2000 American Chemical Society | |||||||||||
