Structure -Function relationship of Membrane Bound Peptides Using DPC Micelles

使用 DPC 胶束的膜结合肽的结构-功能关系

• 批准号: 02453152
• 负责人: INAGAKI Fuyuhiko
• 金额: $ 3.07万
• 依托单位: The Tokyo Metropolitan Institute of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02453152/
• 关键词: Bioactive peptides; Perdeuterated micelles; NMR; Distance geometry; 膜融合性ペプチド; 構造ー機能相関

Biologically active peptides bind to biomembranes and exert several biological actions such as membrane lysis and membrane fusion. These activities are closely related to the structures of the peptides embedded in the membranes, which led us to the detailed investigation of the three dimensional structures of those peptides bound to the membranes. However, the peptides bound to the membranes generally give broad NMR resonances so that we used micelles as model membranes. We synthesized perdeuterated dodecylphosphocholine which made it possible to observe the NMR signals derived from the peptides not from lipids and also reduced the spin diffusion effects. These points are significant advantages of the use of perdeuterated micelles for the structural determination of the membrane bound peptides.We determined the structure of melittin bound to the micelles using NMR and distance geometry calculations. Melittin takes a rod with a bent at Prol4. However, the bent angle can not be determine … More d due to the lack of NOE information. Melittin binds to the surface of membranes with the helical axis parallel to the surface. The flexibility of the rod may be effective for membrane lysis just as wedges destruct objects. We also determined the three dimensional structure of the epidermal growth factor(EGF)bound to the micelles. In this case, the micelles mimic hydrophobic environments of the receptors. EGF binds to the micelles with the C-terminal tail, where C-terminal tail takes an amphiphilic structure. Leu47 has been suggested to be essential for biological activity of EGF. Actually, Leu47 is located at the pivotal position to connect the hydrophobic parts of the Nterminal and C-terminal domains. This amphiphilic structure of EGF is important for the binding of EGF to the receptor. In addition to these studies, we have made the basic investigation on the distance geometry calculations and determined the structure of biologically active peptides such as yi-conotoxin GIIIA and sapecin. Less

生物活性肽与生物膜结合并发挥多种生物作用，例如膜裂解和膜融合。这些活性与嵌入膜中的肽的结构密切相关，这使我们对与膜结合的肽的三维结构进行了详细研究。然而，与膜结合的肽通常会产生广泛的 NMR 共振，因此我们使用胶束作为模型膜。我们合成了全氘化十二烷基磷酸胆碱，这使得可以观察来自肽而非脂质的 NMR 信号，并且还减少了自旋扩散效应。这些点是使用全氘化胶束来确定膜结合肽的结构的显着优势。我们使用核磁共振和距离几何计算确定了与胶束结合的蜂毒肽的结构。 Melittin 在 Prol4 处拿了一根弯曲的棒。然而，由于缺乏 NOE 信息，无法确定弯曲角度。蜂毒肽与膜表面结合，螺旋轴平行于表面。杆的灵活性可能对膜裂解有效，就像楔子破坏物体一样。我们还确定了与胶束结合的表皮生长因子（EGF）的三维结构。在这种情况下，胶束模拟受体的疏水环境。 EGF 通过 C 端尾部与胶束结合，其中 C 端尾部采用两亲结构。 Leu47 被认为对于 EGF 的生物活性至关重要。实际上，Leu47 位于连接 N 端和 C 端结构域疏水部分的关键位置。 EGF 的这种两亲结构对于 EGF 与受体的结合很重要。除了这些研究之外，我们还对距离几何计算进行了基础研究，并确定了yi-芋螺毒素GIIIA和sapecin等生物活性肽的结构。较少的

项目成果

D.Kohda: "Characterization of pH Titration Shifts for All the Nonlabile Proton Resonances in a Protein by Two-Dimensional NMR:The Case of Mouse Epidermal Growth Factor," Biochemistry. 30. 4896-4900 (1991)

D.Kohda：“通过二维 NMR 表征蛋白质中所有不稳定质子共振的 pH 滴定变化：小鼠表皮生长因子的案例”，《生物化学》。

T.Ikura: "Refined Structure of Melittin Bound to Perdeuterated Dodecylphosphocholine Micelles as Studied by 2D-NMR and Distance Geometry Calculation." Proteins:Structure,Function and Genetics. 9. 81-89 (1991)

T.Ikura：“通过 2D-NMR 和距离几何计算研究了与全氘化十二烷基磷酸胆碱胶束结合的蜂毒肽的精细结构。”

D.Kohda,F.Inagaki: "Structure of mouse EGF bound to micelles"

D.Kohda、F.Inagaki：“小鼠 EGF 与胶束结合的结构”

F.Inagaki,K.Kawaguchi: "Structureーfunction relationship of LHRH and its analogs"

F.Inagaki、K.Kawaguchi：“LHRH 及其类似物的结构-功能关系”

J-M. Lancelin, D. Kohda, S. Tate, Y. Yanagawa, T. Abe, M. Satake, and F. Inagaki: "Tertiary Structure of Conotoxin GIIIA in Aqueous Solution" Biochemistry. 30. 6908-6916 (1991)

J-M。