Development of the new method for dynamics analysis in large molecular protein-protein complex using nuclear magnetic resonance

• Project/Area Number: 25840021
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Nagoya University
• Principal Investigator: MIYANOIRI Yohei 名古屋大学, 理学(系)研究科(研究院), 特任助教 (80547521)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: NMR / 蛋白質動態

Outline of Final Research Achievements

To elucidate the dynamic properties of GroEL-ES protein complexs, We have developed new relaxation optimized Stereo-Array Isotope labelling (SAIL) method. Using new relaxation optimized SAIL amino acids, we could succeed in observing well-separeted aromatic, aliphatic and methyl CH signals for GroEL-ES complex. And we could detect the changes in structural flactuations in several amino acid s in GroEL and GroES upon forming GroEL-ES complex. These dynamics may play an important role for cotrolling a biological function of GroEL-ES complex.
Our new SAIL NMR method will be useful for structural dynamics study in many macromolecular protein complexes.

Research Products

• [Journal Article] Expression and purification of a GRAS domain of SLR1, the rice DELLAprotein (2014)
  • Author(s): Sato, T., Miyanoiri, Y., Takeda, M., Naoe, Y., Mitani, R., Hirano, K., Takehara, S., Kainosho, M., Matsuoka, M., Ueguchi-Tanaka, M., Kato, H.
  • Journal Title: Protein Expr. Purif. Volume: N/A Pages: 248-254
  • DOI: 10.1016/j.pep.2014.01.006
  • Peer Reviewed / Open Access
• [Journal Article] Use of H/D isotope effects to gather information about hydrogen bonding and hydrogen exchange rates (2014)
  • Author(s): Mitsuhiro Takeda, Yohei Miyanoiri, Tsutomu Terauchi, Chun-Jiun Yang, Masatsune Kainosho
  • Journal Title: J. Magn. Reson. Volume: 241 Pages: 148-154
  • DOI: 10.1016/j.jmr.2013.10.001
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Differential isotope-labeling for Leu and Val residues in a protein by E. coli cellular expression using stereo-specifically methyl labeled amino acids (2013)
  • Author(s): Yohei Miyanoiri, Mitsuhiro Takeda, Kosuke Okuma, Akira M. Ono, Tsutomu Terauchi, Masatsune Kainosho
  • Journal Title: J. Biomol. NMR Volume: 57 Issue: 3 Pages: 237-249
  • DOI: 10.1007/s10858-013-9784-0
  • Peer Reviewed
• [Journal Article] Molecular bases of multimodal regulation of a fungal transient receptor potential (TRP) channel (2013)
  • Author(s): Ihara M,Hamamoto S, Miyanoiri Y, Takeda M, Kainosho M, Yabe I, Uozumi N, Yamashita A
  • Journal Title: Journal of Biological Chemistry Volume: (in press) Issue: 21 Pages: 15303-17
  • DOI: 10.1074/jbc.m112.434795
  • Peer Reviewed
• [Presentation] 高度な安定同位体標識技術を利用した 高分子量蛋白質の立体構造解析法の開発 (2015)
  • Author(s): 宮ノ入洋平、武田光広、楊淳竣、寺内勉、甲斐荘正恒
  • Organizer: 第四回 岐阜構造生物学・医学・論理的創薬研究会 シンポジウム
  • Place of Presentation: 岐阜薬科大学、岐阜県
  • Year and Date: 2015-03-05
  • Invited
• [Presentation] 大腸菌生合成系を利用した新規SAILアミノ酸標識法の開発 (2014)
  • Author(s): 宮ノ入洋平、石田洋二郎、武田光広、寺内勉、井上正順、甲斐荘正恒
  • Organizer: 第53回 NMR討論会
  • Place of Presentation: 大阪大学、 大阪府
  • Year and Date: 2014-11-04 – 2014-11-06
• [Presentation] Structural analysis of large molecular proteins using SAIL method and non-uniform sampling (2014)
  • Author(s): Miyanoiri Y, Takeda M, Yang CJ, Terauchi T, Kainosho M
  • Organizer: IPR seminar “Practical Aspects of in Non-uniform Sampling in Multi-dimensional NMR Spectroscopy & Applications for Biological Systems”
  • Place of Presentation: Nagoya University, Aichi
  • Year and Date: 2014-06-18 – 2014-06-19
• [Presentation] 高分子量蛋白質の立体構造解析に向けた新規SAILアミノ酸標識法の開発 (2013)
  • Author(s): 宮ノ入洋平、武田光広、寺内勉、甲斐荘正恒
  • Organizer: 第52回 NMR討論会
  • Place of Presentation: 石川県立音楽堂 〒920-0856 石川県金沢市昭和町20-1
• [Remarks] 名古屋大学大学院理学研究科附属構造生物学研究センター 甲斐荘グループ 研究業績
  • URL: http://str.bio.nagoya-u.ac.jp:8080/Plone/753265908358/753265908358/78147a76696d7e3e
• [Remarks] 名古屋大学大学院理学研究科生命理学専攻 論文紹介
  • URL: http://www.bio.nagoya-u.ac.jp/paper/23th.html
• [Remarks] 名古屋大学大学院理学研究科附属構造生物学研究センター 甲斐荘グループ 研究業績
  • URL: http://str.bio.nagoya-u.ac.jp:8080/Plone/753265908358/753265908358/78147a76696d7e3e
• [Remarks] 名古屋大学大学院理学研究科生命理学専攻 論文紹介
  • URL: http://www.bio.nagoya-u.ac.jp/paper/index.html