Analysis of the assembly apparatus of photosystem I reaction center complex

光系统I反应中心复合体组装装置分析

• 批准号: 15570037
• 负责人: TAKAHASHI Yuichiro
• 金额: $ 2.37万
• 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-15570037/
• 关键词: photosynthesis; photosystem; assembly; chloroplast; transformation; Ycf4; tag; affinity chromatography; 緑藻クラミドモナス; サブユニット組成; タグ融合タンパク

Chloroplast-encoded Ycf4 is a thylakoidal protein and is essential for the assembly of photosystem I (PSI) reaction center complex. This protein is a part of a large complex that is an assembly apparatus of PSI complex. In this project, the Ycf4-complex was highly purified by two steps affinity chromatography from the thylakoid extracts of the chloroplast transformant of Chlamydomonas reinhardtii in which Ycf4 fused with TAP-tag is expressed. The resultant Ycf4-complex was found to contain Cop2, the retinal binding protein, and MASAP (mitochondrial ATP synthase associated protein) as well as PsaF, one of the PSI subunits and Ycf4. It was found from separate biochemical experiments that all Cop2 present in the thylakoids is co-purified with Ycf4, indicating that this retinal binding protein is an authentic component of the Ycf4-complex. The functional role of Cop2 is not yet known. Absorption and fluorescence emission spectra revealed that the purified Ycf4-complex contains chlorophyll and carotenoid. These pigments may be supplied to an assembly intermediate of PSI complex.

叶绿体编码的Ycf 4是一种类囊体蛋白，是光系统I（PSI）反应中心复合物组装所必需的。该蛋白质是一个大的复合物的一部分，该复合物是PSI复合物的组装装置。本研究采用两步亲和层析法从莱茵衣藻叶绿体类囊体提取物中分离纯化Ycf 4-复合物，并在其中表达了与TAP-标签融合的Ycf 4。发现所得Ycf 4复合物含有Cop 2（视网膜结合蛋白）和MASAP（线粒体ATP合成酶相关蛋白）以及PsaF（PSI亚基之一）和Ycf 4。从单独的生物化学实验中发现，类囊体中存在的所有Cop 2与Ycf 4共纯化，表明这种视网膜结合蛋白是Ycf 4复合物的真实组分。Cop 2的功能作用尚不清楚。吸收光谱和荧光发射光谱表明，纯化的Ycf 4-复合物含有叶绿素和类胡萝卜素。这些颜料可以被供应到PSI复合物的组装中间体。

项目成果

I.Sugimoto, Y.Takahashi: "Evidence that the PsbK polypeptide is associated with the photosystem II core antenna complex CP43"The Journal of Biological Chemistry. 278. 45004-45010 (2003)

I.Sugimoto、Y.Takahashi：“PsbK 多肽与光系统 II 核心天线复合物 CP43 相关的证据”《生物化学杂志》。

Tokutsu R, Teramoto H, Takahashi Y, Ono T, Minagawa J: "The Light-harvesting complex of photosystem I in Chlamydomonas reinhardtii : Protein composition, gene structures and phylogenic implications"Plant and Cell Physiology. (In press).

Tokutsu R、Teramoto H、Takahashi Y、Ono T、Minakawa J：“莱茵衣藻光系统 I 的光捕获复合体：蛋白质组成、基因结构和系统发育影响”植物和细胞生理学。

The light-harvesting complex of photosystem I in Chlamydomonas reinhardtii: protein composition, gene structures and phylogenic implications.

• DOI: 10.1093/pcp/pch013
• 发表时间: 2004-02
• 期刊: Plant & cell physiology
• 影响因子: 4.9
• 作者: Ryutaro Tokutsu;H. Teramoto;Yuichiro Takahashi;T. Ono;J. Minagawa

N.Ohnishi, Y.Takahashi: "Localization and function of small chloroplast-encoded PsbT polypeptide of photosystem II in the green alga Chlamydomonas reinharrdtii"Proceedings of Endocytobiology (Logos Verlag Berlin). VIII(in press).

N.Ohnishi，Y.Takahashi：“绿藻衣藻莱茵衣藻中光系统 II 的小叶绿体编码的 PsbT 多肽的定位和功能”内细胞生物学论文集（Logos Verlag Berlin）。

Establishment of publicly available cDNA material and information resource of Chlamydomonas reinhardtii (Chlorophyta) to facilitate gene function analysis

• DOI: 10.2216/i0031-8884-43-6-722.1
• 发表时间: 2004-11-01
• 期刊: PHYCOLOGIA
• 影响因子: 1.6
• 作者: Asamizu, E;Nakamura, Y;Tabata, S
• 通讯作者: Tabata, S