1998 Fiscal Year Annual Research Report

^<15>N、^<13>C、^2H固体NMRによる繊維・高分子の非晶構造とダイナミクスの解明

Research Project

Project/Area Number	09450357
Research Institution	TOKYO UNIVERSITY OF AGRICULTURE AND TECHNOLOGY
Principal Investigator	朝倉哲郎東京農工大学, 工学部, 教授 (30139208)
Co-Investigator(Kenkyū-buntansha)	出村誠北海道大学, 理学部, 助教授 (70188704)
Keywords	固体NMR / 繊維・高分子の非晶構造 / 繊維・高分子のダイナミクス / NMR化学シフト / 家蚕およびエリ蚕絹フィブロイン / 同位体ラベル絹 / ポリ乳酸 / ケラチン
Research Abstract	本研究の目的は、繊維・高分子の非晶構造解明のための個体NMRの手法を開発し、その非晶構造ならびにダイナミクスを明らかにするとともに、その物性との関連を解明することである。研究対象は、様々な部位を^<13>C,^<15>Nおよび^2Hで同位体ラベルされた家蚕およびエリ蚕絹、^<13>Cラベル毛髪ケラチン、ポリ乳酸等である。研究手法は、角度依存個体NMR、重水素個体ダイナミクスNMR、等方的NMR化学シフトの決定と化学シフト計算、個体スピン拡散NMR、REDOR法等であり、各種測定は、CMX400MHz固体NMR装置を用いた。例えば、安定同位体ラベルエリ蚕絹について、角度依存個体NMR測定によって配向成分の主鎖内部回転角を決定することができた。家蚕絹同様、βシート構造をとる。その配向成分の割合の決定から、延伸初期にAla連鎖部分がβシート構造を形成し、Gly残基の存在する領域は延伸によって引き延ばされると考えられた。エリ蚕絹の配向成分の割合が家蚕と比べかなり少ないこと、Gly残基の配向軸方向の乱れが大きいなどの要因が、二種類の絹の力学的物性の差に反映されるものと結論した。また、重水素ラベル家蚕及びエリ蚕絹の個体重水素NMRから、Ser側鎖には運動状態の異なる二成分が存在し、遅い成分は、Ser側鎖水素基の酸基の関与した水素結合形成に対応した。一方、家蚕及びエリ蚕絹のTyr環の運動は、二成分で解析でき、家蚕絹では速いflip-flop成分が20%、エリ蚕絹では周囲により動ける空間が存在することを示す。これが、二種の絹間の例えば力学的損失正接tanδの値の違いに対応すると考えられた。さらに、40個のタンパク質の原子座標と化学シフトのデータベースに基づいて、^<13>C NMR化学シフトの(Φ,Ψ)マップをアミノ酸残基毎に作成、それを家蚕絹の繊維化前後の各々SilkI,SilkII型構造及びエリ蚕絹の繊維化前後の各々、α-heix,β構造の解明に適用した。その他、個体スピン拡散NMR、REDOR法を新たに絹構造解析に適用するとともに、ケラチン、ポリ乳酸の^<13>CNMR個体構造解析を行い、構造と物質との関連について研究した。以上のように、繊維・高分子の非晶構造解析のための個体NMRの手法を5種類程、開発でき、その非晶構造ならびにダイナミクスの検討をすすめ、その物性との関連を解明することができた。成果の詳細は、投稿中を含め、合わせて、34報にまとめることができた。

Research Products

(31 results)

All Other

All Publications (31 results)

[Publications] T.Asakura,M.Demura,T.Date,N.Miyashita,K.Ogawa,M.P.Williamson: "NMR Study of Silk I Structure of Bombyx mori Silk Fibroin with ^<15>N-and ^<13>C-NMR Chemical Shift Contour Plots" Biopolymers. 41. 193-203 (1997)
[Publications] T.Asakura,M.Iwadate,T.Date,M.Demura and K.Nokihara: "Refinement of the Solution Structure of Neurokinin A(4-10)on the Basis of ^1H NMR Chemical Shift Calculations" Peptide Chemistry 1996. 345-348 (1997)
[Publications] M.Iwadate,T.Asakura and M.P.Williamson: "The structure of the melittin tetramer at differennt temperatures: An NOE-based calculation with chemical shift refinement" European J.of Biochemistry. 257. 479-487 (1998)
[Publications] M.Iwadate,T.Asakura and M.P.Williamson: "Cα and Cβ Carbon13 Chemical Shifts in Proteins from an Empirical Database" J.Biomol.NMR. (in press).
[Publications] J.Kikuchi,T.Asakura,P.A.Loach,P.S.Parkes-Loach,K.Shimada,C.N.Hunter,M.J.Conroy and M.P.Williamson: "A light-harvesting antenna protein retains its folded conformation in the absence of protein-lipid and protein-pigment interactions" Biopolymers. (in press).
[Publications] J.Kikuchi and T.Asakura: "Use of ^<13>C conformation-dependent chemical shifts to elucidate the local structure of a large protein with homologous domains in solution and solid state" J.Biochem.Biophys.Method. (in press).
[Publications] T.Asakura,M.Iwadate,M.Demura and M.P.Williamson: "Structural Analysis of Silk using ^<13>C NMR Chemical Shift Contour Plots," Int.J.Biolog.Macromol. (in press).
[Publications] J.Kikuchi,T.Asakura,K.Hasuda,T.Ito,K.Ohwaku and M.P.Williamson: "An Advantage for Use of Isotope Labeling and NMR Chemical Shifts to Analyze The Structure of Four Homologous IgG-binding Domains of Staphylococcal Protein A" J.Biochem.Biophys.Method. (submitted).
[Publications] Y.Hori,M.Demura,M.Iwadate,T.Niidome,H.Aoyagi and T.Asakura: Biophysical J.(submitted).
[Publications] T,Asakura,M.Minami,R.Shimada,M.Demura,M.Osanai,T.Fujito,M.Imanari,A.S.Ulrich: "^2H-Labelling of Silk-Fibroin Fibers and Their Structural Characterization by Solid State ^2H NMR" Macromolecules. 30. 2429-2435 (1997)
[Publications] M.Demura,M.Minami,T.Asakura,and T.A.Cross: "Structure of Bombyx mori Silk Fibroin Based on Solid State NMR Orientational Constraints and Fiber Diffraction Unit Cell Parameters" J.Am.Chem.Soc.120. 1300-1308 (1998)
[Publications] M.Demura,Y.Yamazaki,T.Asakura and K.Ogawa: "Structure of Uniaxially Aligned ^<13>C Labeled Silk Fibroin Fibers with Solid State ^<13>C-NMR" J.Molecular Structure. 441. 155-163 (1998)
[Publications] T.Asakura,Y.Yamazaki,Koo Wey Seng and M.Demura: "Determination of the Mutual Orientation of the ^<15>N and ^<13>C NMR Chemical Shift Tensors of ^<13>C-^<15>N Double Labeled Model Peptides for Silk Fibroin from the Dipolar-Coupled Powder Patterns" J.Molecular Structure. 446. 179-190 (1998)
[Publications] T.Ito,Y.Maruhashi,M.Demura and T.Asakura: "Carbon-13 Solid State n.m.r. Study on Uniaxially Oriented Poly(L-lactic acid) Films" Polymers. (submitted).
[Publications] T.Asakura and M.Demura: "Structure and Dynamics of Silk Fibroin Studies with ^<13>C,^<15>N and ^2H Solid State NMR" Macromol.Chem.Macromolecular Symp.(in press).
[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi and T.Asakura: "Structure change of keratin protein in human hair by permanent waving treatment" Polymer. 39,16. 3835-3840 (1998)
[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi,H.Matsuno and T.Asakura: "Carbon-13n.m.r.studies of keratin intermediate filament hair" Polymer. 39,4. 1001-1004 (1998)
[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi,H.Matsuno and T.Asakura: "pH Dependence of the Coiled-Coil Structure of Keratin Intermediate Filament in Human Hair by ^<13>C NMR Spectrosroscopy and the Mechanism of Its Disruption" Polymer J.30,2. 125-132 (1998)
[Publications] J.Kikuchi,Y.Mitsui,T.Asakura,K.Hasuda,H.Araki and K.Owaku: "Spectroscopic inverstigation of tertiary fold of staphylococcal protein A to explore its engineering application" Biomaterials. (in press).
[Publications] N.Nishikawa,Y.Horiguchi,Tetsuo Asakura and Iso Ando: "Carbon-13 solid-state n.m.r.study of 13C-enriched human hair keratin" Polymer. 40. 2139-2144 (1999)
[Publications] Y.Hori,M.Demura,T.Asakura,T.Niidome and H.Aoyagi: "Determination of Molecular Orientation in Peptide-Phospholipid Bilayer System using Angular Dependent Solid State ^<31>P,^2H and ^<15>N NMR" Reports on Progress in Polymer Physics in Japan. 40. 609-612 (1997)
[Publications] M.Demura,Y.Ohkawa and T.Asakura: "Side Chain Dynamics of [3,3-^2H_2]Serine Labeled Silk Fibroin using Solid State ^2H NMR" Reports on Progress in Polymer Physics in Japan. 40. 613-616 (1997)
[Publications] A.Aoki and T.Asakura: "Packing Effect on ^<13>C CP/MAS NMR Spectra of Isotactic Polypropylene" Reports on Progress in Polymer Physics in Japan. 40. 475-478 (1997)
[Publications] J.Kikuchi,T.Yamazaki and T.Asakura: "Characterization of Aggregated State of Light-harvesting Antenna Protein in Detergent Micelles" Reports on Progress in Polymer Physics in Japan. (in press).
[Publications] J.Kikuchi, K.Shimada and T.Asakura: "Solid-state NMR Spectroscopic Study of Membrane-bound Proteins in Rhodobacter Sphaeroides" Reports on Progress in Polymer Physics in Japan. (in press).
[Publications] M.Demura,O.Saita,T.Asakura,J.D.van Beek and Beat.H.Meier: "Determination of Torsion Angles of Silk Fibroin by Two-Dimensional Spin Diffusion NMR" Reports on Progress in Polymer Physics in Japan. (in press).
[Publications] M.P.Williamson and T.Asakura: "Protein Chemical Shifts,Methods in Molecular Biology" Protein NMR Techniques Edited by D.G.Reid Humana Press Inc.,Totowa,NJ.60. (1997)
[Publications] T.Asakura,M.Demura and N.Naoki: "Structural Analysis of Oriented Polymers by Solid-State NMR" (Annual Reports on NMR Spectroscopy)(Ed.G.A.Webb)(ACADEMIC PRESS,London). 34. 301-346 (1997)
[Publications] 朝倉哲郎: "繊維構造に関するNMR研究" 繊維学会誌. 53. 300-304 (1997)
[Publications] 朝倉哲郎、出村誠: "シルク研究の新展開" 高分子. 47. 390-393 (1998)
[Publications] I.Ando and T.Asakura Eds.: "Proteins in “Solid State NMR of Polymers"" Elsevier, 1000 (1998)

1998 Fiscal Year Annual Research Report

^<15>N、^<13>C、^2H固体NMRによる繊維・高分子の非晶構造とダイナミクスの解明

Principal Investigator

朝倉 哲郎 東京農工大学, 工学部, 教授 (30139208)

Research Products

[Publications] T.Asakura,M.Demura,T.Date,N.Miyashita,K.Ogawa,M.P.Williamson: "NMR Study of Silk I Structure of Bombyx mori Silk Fibroin with ^<15>N-and ^<13>C-NMR Chemical Shift Contour Plots" Biopolymers. 41. 193-203 (1997)

[Publications] T.Asakura,M.Iwadate,T.Date,M.Demura and K.Nokihara: "Refinement of the Solution Structure of Neurokinin A(4-10)on the Basis of ^1H NMR Chemical Shift Calculations" Peptide Chemistry 1996. 345-348 (1997)

[Publications] M.Iwadate,T.Asakura and M.P.Williamson: "The structure of the melittin tetramer at differennt temperatures: An NOE-based calculation with chemical shift refinement" European J.of Biochemistry. 257. 479-487 (1998)

[Publications] M.Iwadate,T.Asakura and M.P.Williamson: "Cα and Cβ Carbon13 Chemical Shifts in Proteins from an Empirical Database" J.Biomol.NMR. (in press).

[Publications] J.Kikuchi,T.Asakura,P.A.Loach,P.S.Parkes-Loach,K.Shimada,C.N.Hunter,M.J.Conroy and M.P.Williamson: "A light-harvesting antenna protein retains its folded conformation in the absence of protein-lipid and protein-pigment interactions" Biopolymers. (in press).

[Publications] J.Kikuchi and T.Asakura: "Use of ^<13>C conformation-dependent chemical shifts to elucidate the local structure of a large protein with homologous domains in solution and solid state" J.Biochem.Biophys.Method. (in press).

[Publications] T.Asakura,M.Iwadate,M.Demura and M.P.Williamson: "Structural Analysis of Silk using ^<13>C NMR Chemical Shift Contour Plots," Int.J.Biolog.Macromol. (in press).

[Publications] J.Kikuchi,T.Asakura,K.Hasuda,T.Ito,K.Ohwaku and M.P.Williamson: "An Advantage for Use of Isotope Labeling and NMR Chemical Shifts to Analyze The Structure of Four Homologous IgG-binding Domains of Staphylococcal Protein A" J.Biochem.Biophys.Method. (submitted).

[Publications] Y.Hori,M.Demura,M.Iwadate,T.Niidome,H.Aoyagi and T.Asakura: Biophysical J.(submitted).

[Publications] T,Asakura,M.Minami,R.Shimada,M.Demura,M.Osanai,T.Fujito,M.Imanari,A.S.Ulrich: "^2H-Labelling of Silk-Fibroin Fibers and Their Structural Characterization by Solid State ^2H NMR" Macromolecules. 30. 2429-2435 (1997)

[Publications] M.Demura,M.Minami,T.Asakura,and T.A.Cross: "Structure of Bombyx mori Silk Fibroin Based on Solid State NMR Orientational Constraints and Fiber Diffraction Unit Cell Parameters" J.Am.Chem.Soc.120. 1300-1308 (1998)

[Publications] M.Demura,Y.Yamazaki,T.Asakura and K.Ogawa: "Structure of Uniaxially Aligned ^<13>C Labeled Silk Fibroin Fibers with Solid State ^<13>C-NMR" J.Molecular Structure. 441. 155-163 (1998)

[Publications] T.Ito,Y.Maruhashi,M.Demura and T.Asakura: "Carbon-13 Solid State n.m.r. Study on Uniaxially Oriented Poly(L-lactic acid) Films" Polymers. (submitted).

[Publications] T.Asakura and M.Demura: "Structure and Dynamics of Silk Fibroin Studies with ^<13>C,^<15>N and ^2H Solid State NMR" Macromol.Chem.Macromolecular Symp.(in press).

[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi and T.Asakura: "Structure change of keratin protein in human hair by permanent waving treatment" Polymer. 39,16. 3835-3840 (1998)

[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi,H.Matsuno and T.Asakura: "Carbon-13n.m.r.studies of keratin intermediate filament hair" Polymer. 39,4. 1001-1004 (1998)

[Publications] N.Nishikawa,Y.Tanizawa,S.Tanaka,Y.Horiguchi,H.Matsuno and T.Asakura: "pH Dependence of the Coiled-Coil Structure of Keratin Intermediate Filament in Human Hair by ^<13>C NMR Spectrosroscopy and the Mechanism of Its Disruption" Polymer J.30,2. 125-132 (1998)

[Publications] J.Kikuchi,Y.Mitsui,T.Asakura,K.Hasuda,H.Araki and K.Owaku: "Spectroscopic inverstigation of tertiary fold of staphylococcal protein A to explore its engineering application" Biomaterials. (in press).

[Publications] N.Nishikawa,Y.Horiguchi,Tetsuo Asakura and Iso Ando: "Carbon-13 solid-state n.m.r.study of 13C-enriched human hair keratin" Polymer. 40. 2139-2144 (1999)

[Publications] Y.Hori,M.Demura,T.Asakura,T.Niidome and H.Aoyagi: "Determination of Molecular Orientation in Peptide-Phospholipid Bilayer System using Angular Dependent Solid State ^<31>P,^2H and ^<15>N NMR" Reports on Progress in Polymer Physics in Japan. 40. 609-612 (1997)

[Publications] M.Demura,Y.Ohkawa and T.Asakura: "Side Chain Dynamics of [3,3-^2H_2]Serine Labeled Silk Fibroin using Solid State ^2H NMR" Reports on Progress in Polymer Physics in Japan. 40. 613-616 (1997)

[Publications] A.Aoki and T.Asakura: "Packing Effect on ^<13>C CP/MAS NMR Spectra of Isotactic Polypropylene" Reports on Progress in Polymer Physics in Japan. 40. 475-478 (1997)

[Publications] J.Kikuchi,T.Yamazaki and T.Asakura: "Characterization of Aggregated State of Light-harvesting Antenna Protein in Detergent Micelles" Reports on Progress in Polymer Physics in Japan. (in press).

[Publications] J.Kikuchi, K.Shimada and T.Asakura: "Solid-state NMR Spectroscopic Study of Membrane-bound Proteins in Rhodobacter Sphaeroides" Reports on Progress in Polymer Physics in Japan. (in press).

[Publications] M.Demura,O.Saita,T.Asakura,J.D.van Beek and Beat.H.Meier: "Determination of Torsion Angles of Silk Fibroin by Two-Dimensional Spin Diffusion NMR" Reports on Progress in Polymer Physics in Japan. (in press).

[Publications] M.P.Williamson and T.Asakura: "Protein Chemical Shifts,Methods in Molecular Biology" Protein NMR Techniques Edited by D.G.Reid Humana Press Inc.,Totowa,NJ.60. (1997)

[Publications] T.Asakura,M.Demura and N.Naoki: "Structural Analysis of Oriented Polymers by Solid-State NMR" (Annual Reports on NMR Spectroscopy)(Ed.G.A.Webb)(ACADEMIC PRESS,London). 34. 301-346 (1997)

[Publications] 朝倉哲郎: "繊維構造に関するNMR研究" 繊維学会誌. 53. 300-304 (1997)

[Publications] 朝倉哲郎、出村誠: "シルク研究の新展開" 高分子. 47. 390-393 (1998)

[Publications] I.Ando and T.Asakura Eds.: "Proteins in “Solid State NMR of Polymers"" Elsevier, 1000 (1998)

朝倉哲郎東京農工大学, 工学部, 教授 (30139208)