Determination of the spatial arrangement of polypeptides in photosystem II by photo-and oxygen radical-induced protein cross-linking.

通过光和氧自由基诱导的蛋白质交联测定光系统 II 中多肽的空间排列。

• 批准号: 15570036
• 负责人: YAMAMOTO Yasusi
• 金额: $ 2.05万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15570036/
• 关键词: Photosynthesis; Photosystem II; Protein cross-linking; Reactive oxygen species; Oxygen-evolving system; Polypeptide; Manganese cluster; Protein photodamage; 分子間架橋; 光ストレス; D1タンパク質; プロテアーゼ; FtsH; 熱ストレス; 酵素発生系

The initial purpose of the present research was to identify unknown polypeptides that are located closely to the reaction-center binding D1 protein in photosystem II by the use of chemical cross-linking and light-induced cross-linking methods. However, during the course of the study, we found that a metalloprotease FtsH is located near the core complex of photosystem II. We then have focused our efforts to clarify the roles of the FtsH protease in photosystem II. The following results were obtained.1.By chemical and photochemical cross-linking experiments, we found that the FtsH protease reacts with the D1 protein. These data as well as the membrane fractionation data strongly suggest that the FtsH protease can recognize the photodamaged D1 protein at the grana thylakoids.2.By moderate heat treatment (40゜C for 30 min) on spinach thylakoids, the D1 protein was cleaved. We found that the FtsH is responsible for the cleavage of the heat-damaged D1 protein.3.Using a cyanobacterial mutant lacking the FtsH protease, we confirmed that the the heat-or light-damaged D1 protein in the mutant cells is not removed, and thereby the growth and oxygen-evolving activity are significantly inhibited.All these results strongly suggest that the FtsH protease is essential for the quality control of photosystem II under light and heat stresses.

本研究的最初目的是通过化学交联和光诱导交联的方法鉴定位于光系统II中反应中心结合D1蛋白附近的未知多肽。然而，在研究过程中，我们发现，金属蛋白酶FtsH位于光系统II的核心复合物附近。然后，我们集中我们的努力，以澄清FtsH蛋白酶在光系统II中的作用。主要研究结果如下：1.通过化学交联和光化学交联实验，发现FtsH蛋白酶与D1蛋白发生了反应。这些数据以及膜分离数据都有力地表明FtsH蛋白酶可以识别光损伤的D1蛋白。2.适度热处理（40 ℃，30 min）菠菜类囊体，D1蛋白被切割。我们发现FtsH蛋白酶负责切割热损伤的D1蛋白。3.使用缺乏FtsH蛋白酶的蓝藻突变体，我们证实突变体细胞中的热或光损伤的D1蛋白没有被去除，因此，生长和氧气-所有这些结果都有力地表明，FtsH蛋白酶在光系统II的质量控制中是必不可少的。光和热应力。

项目成果

Isolation and reactive characterization of oxygen-evolving photosystem II complexes retaining the PsbO, P and Q proteins from Euglena gracilis

细小眼虫中保留 PsbO、P 和 Q 蛋白的释氧光系统 II 复合物的分离和反应表征

• 发表时间: 2004
• 期刊: Plant Cell Physiol. 45
• 作者: Jang;M.H.;Ha;K.;Joo;G.J.;Takamura;N.;S.Ohira;S.Ohira;高村典子;高村典子;T.Suzuki
• 通讯作者: T.Suzuki

Assay of photoinhibition of photosystem II and protease activity

光系统 II 的光抑制和蛋白酶活性的测定

• 发表时间: 2004
• 期刊: Methods in Molecular Biology : Photosynthesis Protocols
• 作者: Makino;W.;Mikami;H.;Katano;N.;Nakagawa;M.;Takamura;N.;Y.Yamamoto;Y.Yamamoto
• 通讯作者: Y.Yamamoto

Quality control of photosystem II under light stress - Turnover of the aggregates of the D1 protein in vivo

光应激下光系统 II 的质量控制 - D1 蛋白体内聚集体的周转

• 发表时间: 2005
• 期刊: Photosynthesis Res. (印刷中)
• 作者: Tanaka;S;Ohsaki;N.;S.Ohira
• 通讯作者: S.Ohira

Pressure equilibrium and jump study on unfolding of 23-kDa protein from spinach photosystem II.

• DOI: 10.1529/biophysj.104.050435
• 发表时间: 2005-02
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Cui-yan Tan;Chun-he Xu;J. Wong;Jian-Ren Shen;Shinsuke Sakuma;Yasusi Yamamoto;R. Lange;C. Balny;K. Ruan

Release and reactive-oxygen-mediated damage of the oxygen-evolving complex subunits of PSII during photoinhibition

• DOI: 10.1093/pcp/pch027
• 发表时间: 2004-02-01
• 期刊: PLANT AND CELL PHYSIOLOGY
• 影响因子: 4.9
• 作者: Henmi, T;Miyao, M;Yamamoto, Y
• 通讯作者: Yamamoto, Y