Dynamic structure, signal recognition and material transport of membrane protein

膜蛋白的动态结构、信号识别与物质运输

• 批准号: 14580629
• 负责人: YAMAGUCHI Satoru
• 金额: $ 2.3万
• 依托单位: Himeji Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580629/
• 关键词: Membrane protein; bacteriorhodopsin; phoborhodopsin; fluctuation; dynamics; 固体高分解能NMR; 固体NMR

Two-dimensional array formation and dynamicsThe NMR spectra of Bacteriorhodopsin mutant W12L, W80L that cannot make two-dimensional crystal shows that the protein-protein interaction and protein-lipid interaction change. The dynamics of transmembrane a-helix changes a frequency 10^2 to 10^4 and in 2D crystal, membrane surface has relatively slow mobility (less than 50 μs).The effect of Glu residue for Bacteriorhodopsin stabilityIt was elucidated that Glu^9,Glu^<74>,Glu^<194> and Glu^<204>, which are located at extracellular side, contribute to protein stability. Since Glu^<194> and Glu^<204> are proton release residues, this result implies a relationship between pump function and dynamics.Structure and dynamics of pharaonis phoborhodopsin (ppR) and transducer pHtrII(1-159)A dynamic structure of ppR reconstituted into lipid bilayer agree well with monomeric bacteriorhodopsin. When ppR bind to transducer pHtrII( 1-159), a NMR signal which is assigned loop vanishes because of the interference between dynamics and proton deccooupling frequency (10^5Hz). On the other hand, from NMR spectra, pHtrII(1-159) has α-helix not only transmembranr region but also cytoplasmic side protruding from membrane surface. When ppR-pHtrII(1-159) complex is formed, it become clear that fluctuation of this protruding helix increase and the contribution to signal transduction was suggested.

不能形成二维晶体的细菌视紫红质突变体W12 L、W80 L的核磁共振谱表明，蛋白质与蛋白质、蛋白质与脂质的相互作用发生了变化。跨膜α-螺旋动力学变化的频率为10^2 ~ 10^4，在二维晶体中，膜表面的迁移率相对较慢（小于50 μs）。Glu残基对细菌视紫红质稳定性的影响阐明了位于细胞外侧的Glu ^9、Glu ^<74>、Glu^<194>和Glu^<204>对蛋白质稳定性的贡献。Pharaonis <194><204>phoborhodopsin（ppR）和pHtrII（1-159）的结构和动力学研究当ppR与pHtrII（1-159）结合时，由于动力学和质子去耦频率（10^5Hz）之间的干扰，指定为环的NMR信号消失。另一方面，从NMR谱中，pHtrII（1-159）不仅具有跨膜区，而且具有从膜表面突出的胞质侧。当形成ppR-pHtrII（1-159）复合物时，很明显，这种突出螺旋的波动增加，并提示对信号转导的贡献。

项目成果

H.Saito: "Dynamic Aspect of Bacteriorhodopsin as Viewed from ^<13>C NMR : Conformational Elucidation, Surface Dynamics and Information Transfer from the Surface to Inner Residues"Spectroscopy. 16. 107-120 (2002)

H.Saito：“从 13 C NMR 观察细菌视紫红质的动态方面：构象阐明、表面动力学和从表面到内部残基的信息传递”光谱学。

T.Arakawa: "Dynamic Structure of pharaonis Phoborhodopsin (Sensory Rhodopsin II) and Complex with a Cognate Truncated Transducer as Revealed by Site-directed ^<13>C Solid-state NMR"FEBS Lett.. 536. 237-240 (2003)

T.Arakawa：“通过定点 ^<13>C 固态 NMR 揭示法老荧光紫红质（感觉视紫红质 II）和具有同源截短传感器的复合物的动态结构”FEBS Lett.. 536. 237-240 (2003)

H.saito, T.Tsuchida, K.Ogawa, T.Arakawa, S.Yamaguchi, S.Tuzi: "Residue-specific millisecond to microsecond fluctuations in bacteriorhodopsin induced by disrupted or disorganized two-dimensional crystalline lattice, through modified lipid-helix and helix-h

H.saito、T.Tsuchida、K.Okawa、T.Arakawa、S.Yamaguchi、S.Tuzi：“通过修饰的脂质螺旋，由破坏或无序的二维晶格诱导细菌视紫红质的残留特异性毫秒至微秒波动

H.Saito, R.Kawaminami, M.Tanio, T.Arakawa, S.Yamaguchi, S.Tuzi: "Dynamic aspect of bacteriorhodopsin as viewed from ^<13>C NMR : conformational elucidation, surface dynamics and information transfer from the surface to inner residues"Spectroscopy. 16. 107

H.Saito、R.Kawaminami、M.Tanio、T.Arakawa、S.Yamaguchi、S.Tuzi：“从 13 C NMR 观察细菌视紫红质的动态：构象阐明、表面动力学和表面信息传递

T.Arakawa, K.Shimono, S.Yamaguchi, S.Tuzi, Y.Sudo, N.Kamo, H.Saito: "Dynamic structure of pharaonis phoborhodopsin (sensory rhodopsin II) and complex with a cognate truncated transducer as revealed by site-directed ^<13>C solid-state NMR"FEBS Lett.. 536.

T.Arakawa、K.Shimono、S.Yamaguchi、S.Tuzi、Y.Sudo、N.Kamo、H.Saito：“法老恐惧紫红质（感觉视紫红质 II）和具有同源截短传感器的复合物的动态结构，如位点揭示的