Mechanism of membrane fusion by fusion protein

融合蛋白的膜融合机制

基本信息

批准号：
11694096
负责人：
NAITO Akira
金额：
$ 2.43万
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：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694096/
关键词：
Influenza virus Solid state NMR Fusion peptide lipid bilayer α-helix Neutron scattering Chemical shifts Biomembrane 膜融合活性蛋白質プロトンポンプメリチン αーヘリックス ^<31>P NMR 磁場配向

项目摘要

In this research project, we have studied to understand the molecular mechanism of viral infectn using influenza fusion peptides. We have particularly focused to determine the structure and orientation of the peptides bund to the membrane. Following results were obtained in this research project.(1) 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 ^<31>P and ^<13>C NMR spectroscopy. Using ^<31>P 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. ^<13>C NMR spectra were observed on the ^<13>C labeled melittin bound to the lipid bilayers. Finally, it was found that melittin adopts a transmembrane α-helix whose average axis is parallel to the bilayer normal.(2) N-terminal fragment of HA2 (1-27) in influenza virus was synthesized and investigated by solid state NMR spectroscopy. It was found that the N-terminal part of the fragment forms α-helix and showed different behavior of interaction with membrane at acidic pH from that at basic pH condition.(3) Transmembrane fragment of M2 protein of influenza virus (M2-TMP) was synthesized and investigated by means of ^<15>N solid state NMR spectroscopy. The results indicate that the α-helical axis is tilting by 33° and 37° from the bilayer normalin the DOPC and DMPC bilayer systems, respectively.(4) Membran insertion behavior of influenza fusion peptide was investigated by a neutron scattering method. The results indicate that the helical axis is tilting by 55° from the bilayer normal and the helix is located around the surface of bilayers.

在这个研究项目中，我们研究了使用流感融合肽来了解病毒感染的分子机制，我们特别关注确定肽束在膜上的结构和方向。(1) ）通过固态^<31>P和研究了与二肉豆蔻酰磷脂酰胆碱（DMPC）双层结合的蜂毒肽的构象和动力学以及脂质双层系统中的磁性取向。 13 C NMR光谱。使用13 P NMR，发现蜂毒肽-DMPC双层系统在高于液晶-凝胶相变温度时形成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13 C标记的蜂毒肽上观察13 C NMR谱，最后发现蜂毒肽采用了结构。 (2)合成了流感病毒HA2(1-27)的N端片段,并通过固态核磁共振波谱分析发现N端部分为跨膜α螺旋。该片段的部分形成α螺旋，在酸性pH条件下与碱性pH条件下表现出不同的与膜相互作用的行为。(3)合成并研究了流感病毒M2蛋白的跨膜片段(M2-TMP) 15 N固态NMR光谱结果表明，DOPC和DMPC双层体系中的α-螺旋轴分别与双层法线倾斜33°和37°。(4)膜插入行为。通过中子散射法对流感融合肽进行了研究，结果表明螺旋轴与双层法线倾斜55°，螺旋位于表面周围。双层。