Structural and functional divergence of rhodopsin super-family

视紫红质超家族的结构和功能差异

基本信息

批准号：
21370070
负责人：
KOUYAMA Tsutomu
金额：
$ 12.23万
依托单位：
Nagoya 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-21370070/
关键词：
結晶構造解析ロドプシンプロトンポンプハロロドプシン視物質古細菌レチナール蛋白質 X線結晶構造解析光駆動プロトンポン塩素イオンポンプ膜タンパク質イカ・ロドプシンバクテリオロドプシンアーキロドプシンイカロドプシンイオンポンプ Gタンパク質共役レセプターレチナールタンパク質

项目摘要

(1) Crystallization of retinal proteins by the membrane fusion method:(1) The chloride ion pump from Natoronomonas pharaonis, pharaonis halorhodopsin(pHR), was crystallized, and its structure was determined at 2. 0 Aresolution.(2) Deltarhodopsin(dR3), a light-driven proton pomp found in the cell membrane of Haloterrigena thermotolerans, was crystallized, and its structure was determined at 2. 7 A resolution.(2) Structural analyses of the photoreaction intermediates of bacteriorhodopsin:(1) The structure of the M intermediate was determined at various pH levels. It was shown that the formation of M intermediate at neutral pH is accompanied by a complete destruction of the paired structure of two glutamates in the proton release channel, while this destruction is incomplete at a weakly acidic pH.(2) Using the Leu93-to-Ala mutant(BR_L93A) of bacteriorhodopsin, the structure of the O intermediate was determined at 2. 3 A resolution(3) Structural analyses of the light-driven chloride ion pu … More mp(pHR):(1) The crystal structures of the O-like blue form and the yellow anion-depleted form of pHR were determined at 1. 8 ? 2. 0 A resolutions. The result allowed us to elucidate the mechanism by which the chloride ion is removed from the active site near the protonated Schiff base of retinal.(2) The crystal structure of an M-like intermediate of the pHR-azide complex was determined at 2. 3 A resolution. It is shown that the cytoplasmic half of helix F undergoes a large structural change during the proton pumping cycle(3) A mutant of pHR(pHR_ΔBC) in which a long loop between transmembrane helices B and C was partially deleted was expressed in E. coli. It was shown that the photocycle of pHR_ΔBC is not influenced by the membrane potential, while the uptake process of chloride ion is much faster than observed in the wild-type pHR.(4) Structural analyses of squid rhodopsin(1) The primary photochemical reaction intermediate(Baso) of squid rhodopsin was determined at 2. 7 A resolution.(2) Light-induced structural changes in octopus rhodopsin(oRh) were investigated by small-angle X-ray scattering method. Less

(1)膜融合法视网膜蛋白的结晶：(1)结晶来自Natoronomonas pharaonis的氯离子泵，pharaonis halorhodopsin(pHR)，并在2. 0分辨率下确定其结构。(2)Deltahodopsin(dR3) ），一种在耐热性 Haloterrigena 细胞膜中发现的光驱动质子现象， (2)细菌视紫红质光反应中间体的结构分析:(1)在不同pH值下测定了M中间体的结构表明M的形成。在中性pH下，中间体伴随着质子释放通道中两个谷氨酸的配对结构的完全破坏，而在弱酸性pH下，这种破坏是不完全的。(2)细菌视紫红质的 Leu93-to-Ala 突变体 (BR_L93A)，O 中间体的结构在 2. 3 A 分辨率下测定(3) 光驱动氯离子 pu 的结构分析 mp(pHR):(1)类似 O 的蓝色形式和黄色阴离子耗尽形式的 pHR 的晶体结构以 1. 8 ? 2. 0 A 分辨率测定。结果使我们能够阐明氯离子从视网膜质子化席夫碱附近的活性位点去除的机制。(2) 在 2. 3 处确定了 pHR-叠氮化物复合物的 M 样中间体的晶体结构分辨率表明，螺旋 F 的细胞质一半在质子泵循环期间经历了较大的结构变化(3) pHR(pHR_ΔBC) 的突变体，其中之间存在长环。部分缺失的跨膜螺旋B和C在大肠杆菌中表达，结果表明pHR_ΔBC的光循环不受膜电位的影响，而氯离子的摄取过程比野生型pHR中观察到的要快得多。 (4)鱿鱼视紫红质的结构分析(1)在2. 7 A分辨率下测定了鱿鱼视紫红质的主要光化学反应中间体(Baso)。(2)光诱导采用小角X射线散射法研究了章鱼视紫红质(oRh)的结构变化。