Development of systematic approach in determining three-dimensional structure of membrane bound biomolecules based on accurate interatomic distances

开发基于精确原子间距离确定膜结合生物分子三维结构的系统方法

• 批准号: 09558094
• 负责人: NAITO Akira
• 金额: $ 7.68万
• 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 (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09558094/
• 关键词: Interatomic distance; Dipolar interaction; Three-dimensional structure; REDOR; Molecular dynamics; Membrane bound biomolecule; Isotope labeling; Molecular packing; 原子間距離測定; 回転エコー二重共鳴法; エンケファリン; 化学シフト値; コンフォメーションマップ; リン脂質; 二面体角; 回転エコー二重共鳴

In this research project, we have studied to establish systematic approach in determining the three-dimensional structure of membrane bound biomolecules based on accurate interatomic distances obtained from solid state NMR. Following results were obtained in this research project.(1) The three-dimensional structure of [ィイD113ィエD1C, ィイD115ィエD1N]-labeled Leu-enkephalin (tyr-Gly-Gly-Phe-Leu) dihydrate crystals was determined on the basis of six sets of accurately determined ィイD113ィエD1C-ィイD115ィエD1N interatomic distances by rotational echo double resonance (REDOR) and some additional constraints from ィイD113ィエD1C chemical shifts.(2) The phenyl ring dynamics of [ィイD12ィエD1HィイD25ィエD2]PheィイD14ィエD1-labeled LeuィイD15ィエD1- and MetィイD15ィエD1-enkephalin molecules in crystals grown from four solvents were examined using solid state ィイD12ィエD1H NMR spectroscopy. ィイD12ィエD1H NMR powder pattern clearly indicated the presence of 180°flip motions about the Cβ-Cγbond axis of the phenyl rings. The difference of … More the frequencies for the motion was attributed to the manner of their molecular packing in the crystal.(3) Conformational transition of human calcitonin (hCT) during fibril formation in the acidic and neutral condition were investigated by high resolution solid state ィイD113ィエD1C NMR spectroscopy. The results indicate that conformational transitions from α-helix to β-sheet, and from random coil to β-sheet forms occurred in the central and C-terminus regions, respectively, during fibril formation.(4) The conformation and dynamics of melittin bound to the dimyristoylphophatidylcholin (DMPC) bilayer and the magnetic orientation in the lipid bilayer systems were investigated by solid-state ィイD131ィエD1P and ィイD113ィエD1C NMR spectroscopy. Using ィイD131ィエD1P NMR, it was found that melittin-DMPC bilayer system forms magnetically oriented elongated vesicles with the long axis parallel to the magnetic field above the liquid crystalline-gel phase transition temperature. ィイD113ィエD1C NMR spectra were observed on the ィイD113ィエD1C labeled melittin bound to the lipid bilayer. Finally, it was found that melittin adopts a transmembrane α-helix whose average axis is parallel to the bilayers normal. Less

在这个研究项目中，我们研究了建立系统的方法，在确定膜结合的生物分子的三维结构的基础上，从固态NMR获得精确的原子间距离。本研究取得了以下成果。(1)用旋转回波双共振（REDOR）方法精确测定了六组[酪氨酸D113]-[甘氨酸D115]-[甘氨酸D11N]-亮氨酸脑啡肽（tyr-Gly-Gly-Phe-Leu）二水合物晶体的原子间距，并结合酪氨酸D113]-[甘氨酸D11C]-[甘氨酸D115]-[甘氨酸D1N]-[甘氨酸D1C]-[甘氨酸D1N]-[甘氨酸D1N]-[亮氨酸D1N]-[甘氨酸D1C]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1C]-[甘氨酸D1N]-[甘氨酸D1N]-[亮氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1C]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1C]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1N]-[甘氨酸D1 (2)用固体核磁共振方法研究了[苯丙氨酸D12] D1H-1H-1 NMR谱图表明苯环上的C β-C γ键轴存在180 °翻转运动。的差异 ...更多信息 运动的频率归因于它们在晶体中的分子堆积方式。(3)用高分辨固体核磁共振技术研究了人降钙素（hCT）在酸性和中性条件下纤维形成过程中的构象变化。结果表明，在原纤维形成过程中，α-螺旋向β-折叠和无规卷曲向β-折叠的构象转变分别发生在中心和C-末端区域。(4)用固体核磁共振技术研究了蜂毒肽与二肉豆蔻酰磷脂酰胆碱（DMPC）双层膜结合的构象和动力学以及在脂双层系统中的磁取向。利用双波长D131核磁共振（NMR）研究发现，在液晶-凝胶相变温度以上，蜂毒肽-DMPC双层膜体系形成了长轴平行于磁场的磁性取向的细长囊泡。在与脂质双层结合的蜂毒肽标记的蜂毒肽上观察到了β-D113 β-D1C NMR谱。最后发现蜂毒肽是一种跨膜α-螺旋，其平均轴平行于双层膜法线。少

项目成果

A.Gil: "A ィイD113ィエD1C NMR Study on the Conformation and Dynamical Properties of a Cereal Strorage Protein, C-Hordein, and Its Model Peptides"Biopolymers. 41. 289-300 (1997)

A. Gil：“谷物储存蛋白、C-大麦醇溶蛋白及其模型肽的构象和动力学特性的 D113 D1C NMR 研究”生物聚合物 41. 289-300 (1997)。

S.Tuzi: "Location of a cation-binding site in the loop between helices F and G of bacteriorhodopsin as studied by ^<13>C NMR"Biophys.J.. 76. 1523-1531 (1999)

S.Tuzi：“通过 13 C NMR 研究细菌视紫红质螺旋 F 和 G 之间环中阳离子结合位点的位置”Biophys.J.. 76. 1523-1531 (1999)

M.Tanio: "Conformational changes of bacteriorhodopsin along the proton-conduction chain as studied with ^<13>C NMR of [3-^<13>C]Ala-labeled protein : Arg^<82> may function as an information mediator"Biophys.J.. 77. 1577-1584 (1999)

M.Tanio：“利用 [3-^13C]Ala 标记蛋白的 ^13C NMR 研究细菌视紫红质沿质子传导链的构象变化：Arg^<82> 可能充当信息介体

A.Naito: "Backbone dynamics of polycrystaline peptides studied by measurements of 15N NMR lineshapes and 13C transverse relaxation times" J.Mol.Struct.441. 231-242 (1998)

A.Naito：“通过 15N NMR 线形和 13C 横向弛豫时间的测量研究多晶肽的骨架动力学”J.Mol.Struct.441。

S. Kimura: "A ^<13>C NMR Study on[3-^<13>C]-, [1-^<13>C]Ala or [1-^<13>C]Val-labeled Transmembrane Peptides of Bacteriorhodopsin in Lipid Bilayers : Insertion, Rigid-body Motions and Local Conformational Fuluctuations at Ambient Temperature"Biopolymers. (

S. Kimura：“[3-^ 13 C]-、[1-^ 13 C]Ala 或 [1-^ 13 C]Val 标记跨膜肽的 ^ 13 C NMR 研究