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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)视网膜蛋白质的膜融合法结晶：（1）对法老浮游单胞菌（Natoronomonas pharaonis）的氯离子泵--法老盐视紫红质（pharaonis halorhodopsin，pHR）进行了结晶，并在2 ℃下测定了其结构。0分辨率。(2)三角视紫质（dR 3）是一种存在于嗜热盐地霉细胞膜中的光驱动质子泵，在2。7决议。(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。结果表明，在中性pH下M中间体的形成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，而这种破坏在弱酸性pH下是不完全的。(2)使用细菌视紫红质的Leu 93-to-Ala突变体（BR_L93A），在2. 3A分辨率（3）光驱动氯离子泵的结构分析 ...更多信息 mp（pHR）：（1）在1.八个？2. 0 A决议。这一结果使我们能够阐明氯离子从视黄醛的质子化席夫碱附近的活性位点上被去除的机理。(2)在2.0 ℃下测定了pHR-叠氮化物复合物的类M中间体的晶体结构。3 A决议。（3）在大肠杆菌中表达了跨膜螺旋B和C之间的长环部分缺失的pHR突变体（pHR_ΔBC）。杆菌结果表明，pHR_ΔBC的光循环不受膜电位的影响，而对氯离子的吸收过程比野生型pHR快得多。(4)鱿鱼视紫红质的结构分析（1）测定了鱿鱼视紫红质在2。7决议。(2)用小角X射线散射方法研究了光诱导章鱼视紫红质（oRh）结构的变化。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Functionally important structural elements of the cyano bacterial clock-related protein Pex.

氰基细菌钟相关蛋白 Pex 的功能重要结构元件。

DOI：
发表时间：
2009
期刊：
Genes to Cells 14
影响因子：
0
作者：
Kurosawa S.;Murakami R.;Onai K. (共著第一著者);Morishita M.;Hasegawa D.;Iwase R.;Uzumaki T.;Hayashi F.;Kitajima-Ihara T.;Sakata S.;Murakami M.;Kouyama T.;Ishiura M.
通讯作者：
Ishiura M.

Crystal structure of the O intermediate of the Leu93→Ala mutant of bacteriorhodopsin

细菌视紫红质Leu93→Ala突变体O中间体的晶体结构