2000 Fiscal Year Annual Research Report

膜融合タンパク質の分子機能

Research Project

Project/Area Number	11694096
Research Institution	Himeji Institute of Technology
Principal Investigator	内藤晶姫路工業大学, 理学部, 助教授 (80172245)
Co-Investigator(Kenkyū-buntansha)	斉藤肇姫路工業大学, 理学部, 教授 (30100150)
Keywords	インフルエンザウイルス / 膜融合タンパク質 / 脂質膜 / α-ヘリックス / 生体膜 / 固体NMR / 中性子散乱 / 化学シフト
Research Abstract	本年度は、融合活性をもつインフルエンザウイルスの膜融合ペプチドフラグメントに焦点を絞り、以下に示すように生体膜中での配向と構造を明らかにする研究を行った。 1.インフルエンザウイルスのHA2におけるN端から27アミノ酸残基からなるフラグメントを合成し脂質膜との相互作用について固体NMRを用いて検討した。この結果、フラグメントを膜と強い相互作用を示し、N端付近ではα-ヘリックスを形成していることが判明した。また、酸性条件と中性条件では膜との相互作用に大きな違いのあることが判明した。 2.インフルエンザウイルスのM2タンパク質の膜貫通部分のフラグメント(M2-TMP)を合成し、このペプチドの膜に対する配向の状態について検討した。このため、機械的に配向した膜にフラグメントを再構成し、配向状態で^<15>N-NMRスペクトルを観測した。このペプチドが膜中でα-ヘリックスを形成している場合、^<15>N化学シフト値を詳しく検討したところ、M2-TMPはDOPC膜中では33度傾いており、DMPC膜中では37度傾いていることが判明した。 3.中性子散乱を用いて、インフルエンザウイルスの融合ペプチドの膜挿入について検討した。この結果、ペプチドのヘリックス軸は膜の垂線から55度傾いて膜の表面部位に挿入しているが、ヘリックス自身は膜表面に位置していることが判明した。

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] A.Naito: "Conformation and dynamics of melittin bound to magnetically oriented lipid bilayers by solid-state ^<31>P and ^<13>C NMR spectroscopy"Biophys.J.. 78. 2405-2417 (2000)
[Publications] M.Kamihira: "Conformational transition and fibrillation mechanism of human calcitonin as studied by high-resolution solid-state ^<13>C NMR"Protein Science. 9. 867-877 (2000)
[Publications] S.Yamaguchi: "Irreversible conformational change of bacterio-opsin induced by binding of retinal during its reconstitution of bacterio-opsin, as studied by ^<13>C NMR"J.Biochem.. 127. 861-869 (2000)
[Publications] S.Nakatsuji: "Novel photo-responsive organic spin systems : preparation and properties of norbornadienes and spiropyrans with TEMPO radical substituents"J.Chem.Soc.,Perkin Trans.. 2. 1969-1975 (2000)
[Publications] H.Kimura: "Determination of N-H bond lengths of ^<15>N-labeled poly (L-alanines) by ^1H CRAMPS NMR"Macromolecules. 18. 6627-6629 (2000)
[Publications] J.Bolze: "Small angle X-ray scattering and ^<31>P NMR studies on the phase behavior of phospholipid bilayered mixed micelles"Chem.Phys.Lett.. 329. 215-220 (2000)
[Publications] Y.Kawase: "Alteration of conformation and dynamics of bacteriorhodopsin induced by protonation of Asp85 and deprotonation of Schiff base as studied by ^<13>C NMR"Biochemistry. 47. 14472-14480 (2000)
[Publications] H.Saito: "Conformation and backbone dynamics of bacteriorhodopsin revealed by ^<13>C NMR"Biochim.Biophys.Acta. 1460. 39-48 (2000)
[Publications] S.Kimura: "A ^<13>C NMR study on [3-^<13>C]-.[1-^<13>C] Val labeled transmembrane peptides of bacteriorhodopsin in lipid bilayers : insertion, rigid-body motions at ambient temperature"Biopolymers. 58. 78-88 (2001)
[Publications] K.Nishimura: "Natural abundance ^<13>C REDOR coupled to a singly ^<15>N-labeled nucleus : simulaneous determination of interatomic distances in crystalline ammonium [^<15>N] L-glutamate monohydrate"J.Mol.Struct.. 560. 29-38 (2001)
[Publications] S.Yamaguchi: "Surface dynamics of Bacteriorhodopsin as revealed by ^<13>C NMR studies on [^<13>C] Ala-labeled protein : Determination of millisecond or microsecond motions in interhelical loops and C-terminal α-helix"J.Biochem.. 129. 373-382 (2001)
[Publications] S.Yamaguchi: "Cytoplasmic surfacee structure of bacteriorhodopsin consisting of interhelical loops and C-terminal a-helices modified by a variety of environmental factors as studied by ^<13>C NMR"Eur.J.Biochem.. (in press). (2001)
[Publications] S.Kimura: "Characterization of α-helix structures in polypeptides, revealed by ^<13>C=O…H-^<15>N hydrogen bond lengths determined by ^<13>C REDOR NMR"J.Mol.Struct.. (in press). (2001)

2000 Fiscal Year Annual Research Report

膜融合タンパク質の分子機能

Principal Investigator

内藤 晶 姫路工業大学, 理学部, 助教授 (80172245)

Research Products

[Publications] A.Naito: "Conformation and dynamics of melittin bound to magnetically oriented lipid bilayers by solid-state ^<31>P and ^<13>C NMR spectroscopy"Biophys.J.. 78. 2405-2417 (2000)

[Publications] M.Kamihira: "Conformational transition and fibrillation mechanism of human calcitonin as studied by high-resolution solid-state ^<13>C NMR"Protein Science. 9. 867-877 (2000)

[Publications] S.Yamaguchi: "Irreversible conformational change of bacterio-opsin induced by binding of retinal during its reconstitution of bacterio-opsin, as studied by ^<13>C NMR"J.Biochem.. 127. 861-869 (2000)

[Publications] S.Nakatsuji: "Novel photo-responsive organic spin systems : preparation and properties of norbornadienes and spiropyrans with TEMPO radical substituents"J.Chem.Soc.,Perkin Trans.. 2. 1969-1975 (2000)

[Publications] H.Kimura: "Determination of N-H bond lengths of ^<15>N-labeled poly (L-alanines) by ^1H CRAMPS NMR"Macromolecules. 18. 6627-6629 (2000)

[Publications] J.Bolze: "Small angle X-ray scattering and ^<31>P NMR studies on the phase behavior of phospholipid bilayered mixed micelles"Chem.Phys.Lett.. 329. 215-220 (2000)

[Publications] Y.Kawase: "Alteration of conformation and dynamics of bacteriorhodopsin induced by protonation of Asp85 and deprotonation of Schiff base as studied by ^<13>C NMR"Biochemistry. 47. 14472-14480 (2000)

[Publications] H.Saito: "Conformation and backbone dynamics of bacteriorhodopsin revealed by ^<13>C NMR"Biochim.Biophys.Acta. 1460. 39-48 (2000)

[Publications] S.Kimura: "A ^<13>C NMR study on [3-^<13>C]-.[1-^<13>C] Val labeled transmembrane peptides of bacteriorhodopsin in lipid bilayers : insertion, rigid-body motions at ambient temperature"Biopolymers. 58. 78-88 (2001)

[Publications] K.Nishimura: "Natural abundance ^<13>C REDOR coupled to a singly ^<15>N-labeled nucleus : simulaneous determination of interatomic distances in crystalline ammonium [^<15>N] L-glutamate monohydrate"J.Mol.Struct.. 560. 29-38 (2001)

[Publications] S.Yamaguchi: "Surface dynamics of Bacteriorhodopsin as revealed by ^<13>C NMR studies on [^<13>C] Ala-labeled protein : Determination of millisecond or microsecond motions in interhelical loops and C-terminal α-helix"J.Biochem.. 129. 373-382 (2001)

[Publications] S.Yamaguchi: "Cytoplasmic surfacee structure of bacteriorhodopsin consisting of interhelical loops and C-terminal a-helices modified by a variety of environmental factors as studied by ^<13>C NMR"Eur.J.Biochem.. (in press). (2001)

[Publications] S.Kimura: "Characterization of α-helix structures in polypeptides, revealed by ^<13>C=O…H-^<15>N hydrogen bond lengths determined by ^<13>C REDOR NMR"J.Mol.Struct.. (in press). (2001)

内藤晶姫路工業大学, 理学部, 助教授 (80172245)