1999 Fiscal Year Annual Research Report

人工タンパク質の理論設計と合成

Research Project

Project/Area Number	11166243
Research Institution	Kobe University
Principal Investigator	高田彰二神戸大学, 理学部, 講師 (60304086)
Co-Investigator(Kenkyū-buntansha)	田村厚夫神戸大学, 自然科学研究科, 講師 (90273797)
Keywords	タンパク質 / 人工設計 / デザイン / 遺伝子合成 / 3本へリックス束 / Zスコア / 配列空間アニーリング
Research Abstract	本プロジェクトでは、理論モデルに基づいて人工タンパク質を設計し、それを実際に生合成を行う。理論モデルとしてタンパク質骨格の原子NH、C_αH、COをunited atomとしてあらわに表現する一方、側鎖はその重心に一つの球を置いた粗視化モデルを構築した。理論設計は、鋳型構造に対してもっとも適合するアミノ酸配列(σと書く)を探索する。今回は、鋳型構造としてAlbumin binding domainの3本へリックス束構造を選んだ。適合度としてZスコアを使うのが良い。ランダム配列から、配列σを動的変数としてシミュレーテッドアニーリング法を行い、Zスコアが最適になる配列を10個求めた。我々の粗視化モデルに含まれていない側鎖のぶつかり合いを確認するため、AMBER力場による全原子モデルで構造最適化を行って、そのエネルギーが最低のアミノ酸配列を最終的に理論予測配列とした。この配列に相当する遺伝子を合成し、ベクターDNA(pET11)に組み込み大腸菌(BL21(DE3))を形質転換した。現在、大腸菌の大量培養およびタンパク質精製を進めている。

Research Products
(11 results)

All Other

All Publications (11 results)

[Publications] Shoji Takada 他: "Folding dynamics with nonadditive forces: A simulation study of a designed helical protein and a random heteropolymer"Journal of Chemical Physics. ,110. 11616-11629 (1999)
[Publications] Shoji Takada: "Go-ing for Prediction of Protein Folding Mechanisms"Proceedings of National Academy of Science USA. 96,. 11698-11700 (1999)
[Publications] Shoji Takada: "Simulating Protein Folding with Coarse Grained Models"Supplement of Progress of Theoretical Physics. (印刷中). (2000)
[Publications] 高田彰二: "タンパク質フォールディングの物理化学"生物物理. 227. 20-24 (2000)
[Publications] Shohei Koide, 他: "Multi-step denaturation of Borrelia burgdorferi OspA, a protein containing a single-layer β-sheet"Biochemistry. 38. 4757-4767 (1999)
[Publications] Hiroaki Sasakawa 他: "Backbone 1H, 13C, and 15N resonance assignments of Strepromyces subtilisin inhibitor"J. Biomol. NMR. 14. 285-286 (1999)
[Publications] Peter L. Privalov 他: "Comments on the comments, Protein Engineering,"Protein Engineering. 12,. 187 (1999)
[Publications] Hiroaki Sasakawa, 他: "Secondary structures and structural fluctuation in a dimeric protein, Streptomyces subtilisin inhibitor"J. Biochem. 126. 859-865 (1999)
[Publications] Keisuke Tominaga 他: "Spectroscopic application of THz radiation generated by ultrashort pulses-Static far infrared absorption measurements in condensed phases"Advances in Multiphoton Processes and Spectroscopy. (印刷中). ((2000))
[Publications] Shoji Takada: "Site Resolved Landscape Theory of Fast Folding Protein in "Old and New Views of Protein Folding" edited by K. Kuwajima and M. Arai,"Elsevier. 117-124 (1999)
[Publications] Atsuo Tamura: "Cold denaturation and folding/unfolding of a protein at low temperature in "Old and New Views of Protein Folding" edited by K. Kuwajima and M. Arai,"Elsevier. 279-285 (1999)

1999 Fiscal Year Annual Research Report

人工タンパク質の理論設計と合成

Principal Investigator

高田 彰二 神戸大学, 理学部, 講師 (60304086)

Research Products

[Publications] Shoji Takada 他: "Folding dynamics with nonadditive forces: A simulation study of a designed helical protein and a random heteropolymer"Journal of Chemical Physics. ,110. 11616-11629 (1999)

[Publications] Shoji Takada: "Go-ing for Prediction of Protein Folding Mechanisms"Proceedings of National Academy of Science USA. 96,. 11698-11700 (1999)

[Publications] Shoji Takada: "Simulating Protein Folding with Coarse Grained Models"Supplement of Progress of Theoretical Physics. (印刷中). (2000)

[Publications] 高田彰二: "タンパク質フォールディングの物理化学"生物物理. 227. 20-24 (2000)

[Publications] Shohei Koide, 他: "Multi-step denaturation of Borrelia burgdorferi OspA, a protein containing a single-layer β-sheet"Biochemistry. 38. 4757-4767 (1999)

[Publications] Hiroaki Sasakawa 他: "Backbone 1H, 13C, and 15N resonance assignments of Strepromyces subtilisin inhibitor"J. Biomol. NMR. 14. 285-286 (1999)

[Publications] Peter L. Privalov 他: "Comments on the comments, Protein Engineering,"Protein Engineering. 12,. 187 (1999)

[Publications] Hiroaki Sasakawa, 他: "Secondary structures and structural fluctuation in a dimeric protein, Streptomyces subtilisin inhibitor"J. Biochem. 126. 859-865 (1999)

[Publications] Keisuke Tominaga 他: "Spectroscopic application of THz radiation generated by ultrashort pulses-Static far infrared absorption measurements in condensed phases"Advances in Multiphoton Processes and Spectroscopy. (印刷中). ((2000))

[Publications] Shoji Takada: "Site Resolved Landscape Theory of Fast Folding Protein in "Old and New Views of Protein Folding" edited by K. Kuwajima and M. Arai,"Elsevier. 117-124 (1999)

[Publications] Atsuo Tamura: "Cold denaturation and folding/unfolding of a protein at low temperature in "Old and New Views of Protein Folding" edited by K. Kuwajima and M. Arai,"Elsevier. 279-285 (1999)

高田彰二神戸大学, 理学部, 講師 (60304086)