Elucidation of the maturation mechanism of subtilisins from hyperthermophiles and development of their potential use

阐明来自超嗜热菌的枯草杆菌蛋白酶的成熟机制及其潜在用途的开发

• 批准号: 21380065
• 负责人: KANAYA Shigenori
• 金额: $ 11.98万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21380065/
• 关键词: タンパク質工学; 微生物酵素; 構造生物学; サチライシン; セリンプロテアーゼ; 超好熱菌; 成熟化; プロペプチド; プリオン; タンパク工学; ジェリーロール; X線結晶構造解析; セルピン

The maturation mechanisms of subtilisins, Tk-subtilisin and Tk-SP, from the hyperthermophilic archaeon Thermococcus kodakarensis were analyzed. The results indicate that both proteins are activated(matured) upon autoprocessing and degradation of N-terminal propeprides, folding of Tk-subtilisin is induced upon binding of the Ca^<2+> ions to the Ca^<2+>-binding loop, Tk-SP is matured in the absence of the Ca^<2+> ions, Tk-SP requires C-terminalβ-jelly roll domain for hyperstability. It was also shown that Tk-subtilisin is useful for degradation of abnormal prion proteins.

对极端嗜热古菌Thermococcus kodakarensis的枯草杆菌蛋白酶Tk-subtilisin和Tk-SP的成熟机制进行了分析。结果表明，这两种蛋白质都是在N-末端丙虫啉的自动加工和降解后被激活（成熟）的，Tk-枯草杆菌蛋白酶的折叠是在Ca^2+离子与Ca^2+结合环结合后诱导的，Tk-SP在没有Ca^2+离子的情况下成熟，Tk-SP需要C-末端β-果冻卷结构域来实现超稳定性。还显示Tk-枯草杆菌蛋白酶可用于降解异常朊病毒蛋白。

项目成果

Subtilisin-like serine protease from hyperthermophilic archaeon Thermococcus kodakaraensis with N- and C-terminal propeptides

• DOI: 10.1093/protein/gzp092
• 发表时间: 2010-05-01
• 期刊: PROTEIN ENGINEERING DESIGN & SELECTION
• 影响因子: 2.4
• 作者: Foophow, T.;Tanaka, S.;Kanaya, S.
• 通讯作者: Kanaya, S.

Role of the Ca2+ ions for stability of subtilisin homologue, Tk-SP, from Thermococcus kodakaraensis

Ca2 离子对来自 Thermococcus kodakaraensis 的枯草杆菌蛋白酶同系物 Tk-SP 稳定性的作用

• 发表时间: 2011
• 作者: Nitat Sinsereekul;Tita Foophow;Clement Angkawidjaja;Yuichi Koga;Kazufumi Takano;Shigenori Kanaya
• 通讯作者: Shigenori Kanaya

Accelerated maturation of Tk-subtilisin by the mutation at the C-terminus of propeptide

通过前肽 C 端突变加速 Tk-枯草杆菌蛋白酶的成熟

• 发表时间: 2011
• 作者: Ryo Uehara;Yuichi Koga;Kazufumi Takano;Shigenori Kanaya
• 通讯作者: Shigenori Kanaya

Identification of the Interactions Critical for Propeptide-Catalyzed Folding of Tk-Subtilisin

• DOI: 10.1016/j.jmb.2009.09.028
• 发表时间: 2009-11-27
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Tanaka, Shun-ichi;Matsumura, Hiroyoshi;Kanaya, Shigenori
• 通讯作者: Kanaya, Shigenori

Crystal Structure of a Subtilisin Homologue, Tk-SP, from Thermococcus kodakaraensis: Requirement of a C-terminal β-Jelly Roll Domain for Hyperstability

• DOI: 10.1016/j.jmb.2010.05.064
• 发表时间: 2010-07-23
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Foophow, Tita;Tanaka, Shun-ichi;Kanaya, Shigenori
• 通讯作者: Kanaya, Shigenori