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An Escherichia coli expression system for glutamyl endopeptidases optimized by complete suppression of autodegradation
http://hdl.handle.net/10069/19225
http://hdl.handle.net/10069/19225be533a9d-f796-4e80-9d06-cf97d57b0e78
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
AnaBio381_74.pdf (237.2 kB)
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| Item type | 学術雑誌論文 / Journal Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2008-09-02 | |||||
| タイトル | ||||||
| タイトル | An Escherichia coli expression system for glutamyl endopeptidases optimized by complete suppression of autodegradation | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Autoproteolysis | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Glutamyl endopeptidase | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Prosequence | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Staphylococcus warneri | |||||
| キーワード | ||||||
| 主題Scheme | Other | |||||
| 主題 | Staphylococcus epidermidis | |||||
| 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 | |||||
| 著者 |
Ono, Toshio
× Ono, Toshio× Nemoto, Takayuki K× Shimoyama, Yu× Kimura, Shigenobu× Ohara-Nemoto, Yuko |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | V8 protease (GluV8), a member of the glutamyl endopeptidase I family isolated from the V8 strain of Staphylococcus aureus, is widely used for proteome analysis because of its unique substrate specificity and resistance to detergents. We recently developed an Escherichia coli expression system for the production of GluV8 based on a technique that suppresses the autoproteolysis?the use of the prosequence of its homologue (GluSE) from Staphylococcus epidermidis as a chimeric form or the introduction of four substitutions in the prosequence of GluV8. In the current study, we refined this technique through five amino acid substitutions within the prosequence of GluV8 for complete suppression of the autodegradation. As a result, the recovery of GluV8 proform was enhanced to 20 fg/cell, which was comparable to the level of a constitutive inactive form of GluV8, indicating complete suppression of the autoproteolysis. This mutated propeptide was also effective for the expression of the mature sequence of the glutamyl endopeptidase from Staphylococcus warneri. The recombinant proteins were successfully converted to their active forms through a common cleavage mechanism mediated by thermolysin in vitro. This strategy may shed light on the way for the expression of the proteases that have been scarcely produced in E. coli to date. | |||||
| 書誌情報 |
Analytical Biochemistry 巻 381, 号 1, p. 74-80, 発行日 2008-10-01 |
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| 出版者 | ||||||
| 出版者 | Elsevier B.V. | |||||
| ISSN | ||||||
| 収録物識別子タイプ | ISSN | |||||
| 収録物識別子 | 00032697 | |||||
| PubMed番号 | ||||||
| 関連タイプ | isVersionOf | |||||
| 識別子タイプ | PMID | |||||
| 関連識別子 | 18616923 | |||||
| DOI | ||||||
| 関連タイプ | isVersionOf | |||||
| 識別子タイプ | DOI | |||||
| 関連識別子 | 10.1016/j.ab.2008.06.022 | |||||
| 権利 | ||||||
| 権利情報 | Copyright (c) 2008 Elsevier Inc. All rights reserved. | |||||
| 著者版フラグ | ||||||
| 出版タイプ | AM | |||||
| 出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
| 引用 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | Analytical Biochemistry, 381(1), pp.74-80; 2008 | |||||
