Elucidation of Cl--pumping mechanism and significance of trimer formation of halorhodopsin

阐明氯泵泵机制及盐视紫红质三聚体形成的意义

• 批准号: 19390009
• 负责人: KAMO Naoki
• 金额: $ 11.9万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19390009/
• 关键词: ハロロドプシン; センソリーロドプシン; フォボロドプシン; レチナールタンパク; 高度好塩菌; オーサイト; フォトサイクル; 閃光光分解; exciton-coupling; Circular Dichroism; C1輸送; dodecy1-β-D-maltoside; MALDI-TOF; 解離定数; ホトサイクル; 微生物ロドプシン; Natronomonas Pharaonis; halorhodopsin; イオンポンプ; フラッシュフォトリシス; C

Halorhodopsin (HR) is a light-driven Cl^- pump expressed in the membrane of Halobacteria. We succeeded in developing a new method : HR was expressed in Xenopus laevis Oocytes, and photo-induced membrane current was measured due to the Cl^- transport. This method is highly quantitative. We prepared various HR mutants using E. coli expression system. Careful investigation on both the photochemistry and transport activities of the mutants has been done. We found Arg123 that is an essentially important amino acid residue. During the transition from N to O intermediate, Cl is released to cytoplasm while Cl is uptaken from extracellular space during O to HR' transition. We showed that HR forms trimer even in the presence of detergents, and that the trimer is more robust than the monomer.

盐视紫红质（HR）是在盐细菌的膜中表达的光驱动的Cl^-泵。我们成功地开发了一种新的方法：在非洲爪蟾卵母细胞中表达HR，并测量由于Cl^-转运引起的光诱导膜电流。该方法是高度定量的。我们用E. coli表达系统。对突变体的光化学和运输活性进行了仔细的研究。我们发现Arg 123是一个重要的氨基酸残基。在从N到O的过渡过程中，Cl被释放到细胞质中，而在O到HR的过渡过程中，Cl从细胞外空间被吸收。我们表明，HR形成三聚体，即使在洗涤剂的存在下，和三聚体是更强大的比单体。

项目成果

The photochemical reaction cycle and photoinduced proton transfer of sensory rhodopsin II(phoborhodopsin)from Halobacterium salinarum

盐杆菌感觉视紫红质II（phoborhodopsin）的光化学反应循环和光诱导质子转移

• 发表时间: 2010
• 期刊: Biophys.J. 98
• 作者: J.Tamogami;et al
• 通讯作者: et al

The lifetimes of pharaonis phoborhodopsin signaling states depend on the rate of proton transfers-Effects of hydrostatic pressure and stopped flow experiments

法老磷视紫红质信号状态的寿命取决于质子转移的速率-静水压力和停流实验的影响

• 发表时间: 2008
• 期刊: Photochem. Photobiol. 84
• 作者: T. Kikukawa;他7名
• 通讯作者: 他7名

Heterologous expression of Pharaonis halorhodopsin in Xenopus laevis oocytes and electrophysiological characterization of its light-driven C-pump activity

非洲爪蟾卵母细胞中 Pharaonis 盐视紫红质的异源表达及其光驱动 C 泵活性的电生理学表征

• 发表时间: 2007
• 期刊: Biophysical Journal 92
• 作者: A. Seki;他7名
• 通讯作者: 他7名

Effect of molecular assembly on photointermediates of BR reconstituted into dimyristoyolphosphatidylcholine vesicles.

分子组装对 BR 光中间体重构为二肉豆蔻酰磷脂酰胆碱囊泡的影响。

• 发表时间: 2008
• 作者: M. Sonoyama;T. Kikukawa;Y. Yokoyama;M. Demura;N. Kamo;S. Mitaku
• 通讯作者: S. Mitaku

The chloride ion transport from Natronomonas pharaonis: Role of Thr 218 on the chloride releasing to cytoplasm

Natronomonas pharaonis 的氯离子转运：Thr 218 对氯离子释放到细胞质的作用

• 发表时间: 2007
• 作者: Makise;M.;Takehara;M.;Kuniyasu;A.;Matsui;N.;Nakayama;H. and Mizushima;T;M.Kurihara;Takuma Kitanishi;重村 洋明
• 通讯作者: 重村 洋明