Protein Structural Information Analysis Team





核磁気共鳴（NMR）法は生理的条件下における生体高分子の立体構造情報を提供する分光法です。

解析対象タンパク質を含む水溶液試料に任意の結合リガンドを添加し、そのNMRスペクトル変化から定量性のある、原子レベルの相互作用情報を得ることが 可能な測定法であるといえます。
その一方で、高感度・高精度の解析を行うため安定同位体を試料に取りこませ、かつ凝集のない状態で測定しなければならないことや、 解析対象分子量が巨大になると測定が困難になる、といった制約があります。

我々は、NMRによる膜タンパク質等高分子量タンパク質−リガンド複合体の相互作用解析のため、適切な試料調製方法の確立や、測定技術開発、 および創薬標的タンパク質を対象とした実証研究を行うとともに、NMR相互作用解析情報を活用した機能性分子創出技術の開発も進めています。

2011

Rapid identification of protein-protein interfaces for the construction of a complex model based on multiple unassigned signals by using time-sharing NMR measurements. J Struct Biol. 2011 in press. Kodama Y, Reese ML, Shimba N, Ono K, Kanamori E, Dötsch V, Noguchi S, Fukunishi Y, Suzuki EI, Shimada I, Takahashi H.

Ubiquitination of K-Ras enhances activation and facilitates binding to select downstream effectors. Sci Signal. 4, 13 Sasaki AT, Carracedo A, Locasale JW, Anastasiou D, Takeuchi K, Kahoud ER, Haviv S, Asara JM, Pandolfi PP, Cantley LC.

2010

HNCA-TOCSY-CANH experiments with alternate (13)C- (12)C labeling: a set of 3D experiment with unique supra-sequential information for mainchain resonance assignment. J Biomol NMR. 49, 17-26. Takeuchi K, Gal M, Takahashi H, Shimada I, Wagner G.

Precise structural determination of weakly binding peptides by utilizing dihedral angle constraints. J Biomol NMR. 46, 299-305. Mizukoshi Y, Nagasu M, Shimada I, Takahashi H.

Structural basis for platelet antiaggregation by angiotensin II type 1 receptor antagonist losartan (DuP-753) via glycoprotein VI. J Med Chem. 53, 2087-93. Ono K, Ueda H, Yoshizawa Y, Akazawa D, Tanimura R, Shimada I, Takahashi H.

Direct determination of the insulin-insulin receptor interface using transferred cross-saturation experiments. J Med Chem. 53, 1917-22. Nakamura T, Takahashi H, Takahashi M, Shimba N, Suzuki E, Shimada I.

Real-time assay method of lipid extraction activity. Anal Biochem. 399, 162-7. Sugiki T, Takahashi H, Nagasu M, Hanada K, Shimada I.

Production of isotopically labeled heterologous proteins in non-E. coli prokaryotic and eukaryotic cells. J Biomol NMR. 46, 3-10. Takahashi H, Shimada I.

2009

Structural basis of the interaction between chemokine stromal cell-derived factor-1/CXCL12 and its G-protein-coupled receptor CXCR4. J Biol Chem. 284, 35240-50. Kofuku Y, Yoshiura C, Ueda T, Terasawa H, Hirai T, Tominaga S, Hirose M, Maeda Y, Takahashi H, Terashima Y, Matsushima K, Shimada I.

Structural investigation of the interaction between LolA and LolB using NMR. J Biol Chem. 284, 24634-43. Nakada S, Sakakura M, Takahashi H, Okuda S, Tokuda H, Shimada I.

High-throughput screening of optimal solution conditions for structural biological studies by fluorescence correlation spectroscopy. Protein Sci. 18, 1115-20. Sugiki T, Yoshiura C, Kofuku Y, Ueda T, Shimada I, Takahashi H.

Structural and interaction analysis of glycoprotein VI-binding peptide selected from a phage display library. J Biol Chem. 284, 10720-7. Kato-Takagaki K, Mizukoshi Y, Yoshizawa Y, Akazawa D, Torii Y, Ono K, Tanimura R, Shimada I, Takahashi H.

2008

Stable isotope labeling of protein by Kluyveromyces lactis for NMR study. J. Biol. NMR. 42, 159-62. Sugiki T, Shimada I, Takahashi H.

Cross-saturation and transferred cross-saturation experiments. Prog. NMR Spectrosc. 54, 123-140. Shimada I, Ueda T, Matsumoto M, Sakakura M, Osaswa M, Takeuchi K, Nishida N, and Takahashi H.

2007

Identification and characterization of the slowly exchanging pH-dependenct conformational rearrangement in KcsA. J. Biol. Chem. 282, 15179-15186. K. Takeuchi, H. Takahashi, S. Kawano and I. Shimada

Structural basis of the transition-state stabilization in antibody-catalyzed hydrolysis. J. Mol. Biol. 367, 133-147. M. Sakakura, H. Takahashi, N. Shimba, I. Fujii and I. Shimada

Pairwise NMR experiments for the determination of protein backbone dihedral angle Φ based on cross-correlated spin relaxation. J. Biomol. NMR. 37, 179-185. H. Takahashi and I. Shimada

Backbone resonance assignments for the periplasmic chaperone LolA from Escherichia coli. Biomol. NMR Assign. 1, 125-127. S. Nakada, H. Takahashi, M. Sakakura, M. Kurono and I. Shimada

Backbone resonance assignment for the outer membrane lipoprotein receptor LolB from Escherichia coli. Biomol. NMR Assign. 1, 121-123. S. Nakada, M. Sakakura, H. Takahashi, H. Tokuda and I. Shimada

2006

Utilization of methyl proton resonances in cross-saturation measurement for determining the interface of large protein-protein complexes. J. Biomol. NMR. 34, 167-177. H. Takahashi, M. Miyazawa, Y. Ina, Y. Fukunishi, Y. Mizukoshi, H. Nakamura and I. Shimada

Rapid preparation of stable isotope labeled peptides that bind to target proteins by phage library system. J. Biomol. NMR. 34, 23-30. Y. Mizukoshi, H. Takahashi and I. Shimada

2005

Direct determination of a membrane-peptide interface using the nuclear magnetic resonance cross-saturation method. Biophys. J. 89 4051-4055. T. Nakamura, H. Takahashi, K. Takeuchi, T. Kohno, K. Wakamatsu and I. Shimada

Backbone Resonance Assignment for the Fv Fragment of Catalytic Antibody 6D9 Complexed with a Transition State Analogue. J. Biomol. NMR. 33, 282. M. Sakakura, H. Takahashi, H. Terasawa, K. Takeuchi, I. Fujii and I. Shimada

Conformational dynamics of complementary determining region H3 of an anti-dansyl Fv fragment in the presence of its hapten. J. Mol. Biol. 351 627-640. M. Nakasako, T. Oka, M. Masumo, H. Takahashi, I. Shimada, Y. Yamaguchi, K. Kato and Y. Arata

2004

Channel-forming membrane permeabilization by an antibacterial protein, sapecin: determination of membrane-buried and oligomerization surfaces by NMR. Biochem. J. 279, 4981-4987. K. Takeuchi, H. Takahashi, M. Sugai, H. Iwai, T. Kohno, K. Sekimizu, S. Natori and I. Shimada

Molecular basis of the high-affinity activation of type 1 ryanodine receptors by imperatoxin A. J. Biol. Chem. 377, 385-394. C. W. Lee, E. H. Lee, K. Takeuchi, H. Takahashi, I. Shimada, K. Sato, S. Y. Shin, D. H. Kim and J. I. Kim

2003

Structural basis of the KcsA K+ channel and agitoxin2 pore-blocking toxin interaction by using the transferred cross-saturation method. Structure. 11, 1381-1392. K. Takeuchi, M. Yokogawa, T. Matsuda, M. Sugai, S. Kawano, T. Kohno, H. Nakamura, H. Takahashi and I. Shimada

Novel collagen-binding mode of the VWA domain determined by a transferred cross-saturation experiment. Nat. Struct. Biol. 10, 53-58. N. Nishida, H. Sumikawa, M. Sakakura, N. Shimba, H. Takahashi, H. Terasawa, E. Suzuki and I. Shimada

2002

Backbone 1H, 13C, 15N resonance assignments of the von Willebrand factor A3 domain. J. Biomol. NMR. 24, 357-358. N. Nishida, M. Miyazawa, H. Sumikawa, M. Sakakura, N. Shimba, H. Takahashi, H. Terasawa, E. Suzuki and I. Shimada

Solution structure of ω-grammotoxin SIA, a gating modifier of P/Q and N-type Ca2+ channel. J. Mol. Biol. 321, 517-526. K. Takeuchi, E. J. Park, C. W. Lee, J. I. Kim, H. Takahashi, K. J. Swartz and I. Shimada

Determination of the interface of a large protein complex by transferred cross-saturation measurements. J. Mol. Biol. 318, 245-249. T. Nakanishi, M. Miyazawa, M. Sakakura, H. Terasawa, H. Takahashi and I. Shimada

重点課題（1）-a　極低温電子顕微鏡による高分解能構造解析へ
重点課題（1）-b　Ｘ線結晶構造解析による複合体の構造へ
重点課題（3）　理論計算による研究へ