2000 Fiscal Year Annual Research Report

自発的磁場配向膜を用いた膜機能性生体分子の精密立体構造解析

Research Project

Project/Area Number	11440208
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)	辻暁姫路工業大学, 理学部, 助手 (60227387) 斉藤肇姫路工業大学, 理学部, 教授 (30100150)
Keywords	磁場配向膜 / 膜機能性生体分子 / 長楕円型小胞体 / 脂質二重膜 / 化学シフトテンソル / 膜融合 / 同位体標識 / 微分干渉顕微鏡
Research Abstract	本年度は自発的磁場配向膜に結合した膜機能性生体分子の膜に対する配向および立体構造を決定し、膜機能性生体分子の構造と機能の相関を分子論的に解明する研究を行い以下の成果を得た。 1.これまで明らかにしてきた長楕円型小胞体を形成するメリチン-DMPC小胞体の膜分断における形態変化を微分干渉光学顕微鏡を用いて観測した。この結果、脂質二重膜は相転移点(Tm)以上の温度では、膜融合を繰り返し30μmにも達する巨大小胞体を形成することが判明した。さらに(Tm)付近に温度を下げることで膜表面に小孔を形成して、引き続き膜分断の起こることが明らかとなった。メリチンが膜に小孔を形成することはこれまでも予想されていたが、実際にその像を観測したのは今回が始めてである。 2.自発的磁場配向膜に結合したメリチンの動的な膜結合構造と膜に対する配向の情報を得るため、メリチンのカルボニル炭素を^<13>C同位体標識した試料を合成し、自発的磁場配向膜に組み込んだ。このカルボニル炭素の化学シフトテンソルを静止条件(磁場配向条件)、低速MAS条件、および高速MAS条件で測定した。この結果、カルボニル炭素の^<13>C等方化学シフト値から、メリチンは膜中でN端およびC端に渡ってα-ヘリックス構造をとっていることが判明した。また低速MASによる化学シフト異方性を観測した結果、ヘリックス回りの速い運動の存在が明らかになった。 3.磁場配向状態のメリチンの化学シフト値の解析から、メリチンのヘリックスはヘリックス軸から傾いた軸の回りで歳差運動をしており、その回転軸は膜面に垂直であることが判明した。

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

自発的磁場配向膜を用いた膜機能性生体分子の精密立体構造解析

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)