ログイン
言語:

WEKO3

  • トップ
  • ランキング
To
lat lon distance
To

Field does not validate



インデックスリンク

インデックスツリー

メールアドレスを入力してください。

WEKO

One fine body…

WEKO

One fine body…

アイテム

  1. 010 学術雑誌論文
  2. 桑島, 邦博 / KUWAJIMA, Kunihiro

Folding of Green Fluorescent Protein and the Cycle3 Mutant

https://ir.soken.ac.jp/records/4296
https://ir.soken.ac.jp/records/4296
89755c3d-65e6-4f94-86c4-44582b29a33b
Item type 学術雑誌論文 / Journal Article(1)
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

FUKUDA, Hiroyuki

Search repository
ARAI, Munehito

× ARAI, Munehito

ARAI, Munehito

Search repository
KUWAJIMA, Kunihiro

× KUWAJIMA, Kunihiro

KUWAJIMA, Kunihiro

Search repository
著者別名 桑島, 邦博

× 桑島, 邦博

桑島, 邦博

Search repository
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
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
戻る
0
views
See details
Views

Versions

Ver.1 2023-06-20 14:25:32.391358
Show All versions

Share

Mendeley Twitter Facebook Print Addthis

Cite as

エクスポート

OAI-PMH
  • OAI-PMH JPCOAR 2.0
  • OAI-PMH JPCOAR 1.0
  • OAI-PMH DublinCore
  • OAI-PMH DDI
Other Formats
  • JSON
  • BIBTEX

Confirm


Powered by WEKO3


Powered by WEKO3