Autoprocessing mechanism of SARS-CoV 3CL protease

SARS-CoV 3CL蛋白酶的自动加工机制

• 批准号: 20570115
• 负责人: MURAMATSU Tomonari
• 金额: $ 2.58万
• 依托单位: The Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20570115/
• 关键词: SARS; プロテアーゼ; プロセシング; 基質認識; 基質特異性; 立体構造; 特異性; X線結晶構造解析

3C like protease (3CLPro) of severe acute respiratory syndrome coronavirus (SARS-CoV) is synthesized as a part of polyprotein, and is excised by its own proteolytic activity. It has been previously reported that the precise processing of N-terminus of 3CLPro is required for efficient peptidase activity toward the florogenic peptide substrate. However, by the use of an E. coli in vitro protein synthesis system, we found that the C-terminal site of 3CLPro was efficiently cleaved even though N-terminal pro-sequence was not removed. Moreover, the substrate specificity of this autoprocessing activity was different from that of the mature enzyme. We also found the pocket for this special recognition by X-ray crystallography.

严重急性呼吸综合征冠状病毒（SARS-CoV）的3C样蛋白酶（3CLPro）是由多聚蛋白的一部分合成，并通过自身的蛋白水解作用被切除。据报道，3CLPro的N-末端的精确加工是高效的肽酶活性所必需的。然而，通过使用E. coli体外蛋白质合成系统中，我们发现3CLPro的C-末端位点被有效地切割，即使N-末端pro-sequence没有被去除。此外，这种自加工活性的底物特异性不同于成熟酶。我们还通过X射线晶体学发现了这种特殊识别的口袋。

项目成果

レトロウイルスのがん遺伝子は細胞起源(菊池洋 編)(ノーベル賞の生命科学入門 RNAが拓く新世界 第4章)

逆转录病毒癌基因起源于细胞（菊池博主编）（诺贝尔奖生命科学概论，RNA打开的新世界，第4章）

• 发表时间: 2009
• 作者: 今井瑞依;井上郁;野本直子;内田毅;新澤(伊藤)恭子;吉川信也;石森浩一郎;村松知成
• 通讯作者: 村松知成

The effect of Mutations in the Carboxyl-Terminal Region on the Catalytic Activity of Escherichia coli Signal Peptidase I

羧基端区域突变对大肠杆菌信号肽酶I催化活性的影响

• 发表时间: 2008
• 期刊: J.Biochem 143
• 作者: Kim;Y.-T.;Yoshida;H.;Kojima;M.;Kurita;R.;Nishii;W.;Muramatsu;T.;Ito;H.;Park S.-J.;Takahashi;K.
• 通讯作者: K.