Protein NMR method developed for macromolecular complex

• Project/Area Number: 26650018
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Osaka University
• Principal Investigator: Kojima Chojiro 大阪大学, たんぱく質研究所, 准教授 (50333563)
• Co-Investigator(Renkei-kenkyūsha): FUJIWARA Toshimichi 大阪大学, 蛋白質研究所, 教授 (20242381) ; MATSUKI Yoh 大阪大学, 蛋白質研究所, 助教 (70551498) ; SUGIKI Toshihiko 大阪大学, 蛋白質研究所, 助教 (70635698) ; FURUITA Kyoko 大阪大学, 蛋白質研究所, 研究員 (30727665)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: NMR / 蛋白質 / 立体構造

Outline of Final Research Achievements

Protein structure can be determined by solution NMR precisely, however, molecular weight is limited to below 100kDa. In this study, NMR method has been developed for protein structure analysis, especially for very big protein beyond 100kDa. Here, double-acquisition method was developed to acquire twice data within 1 scan. This double-acquisition method was combined with 13C direct detection method, and optimized for macromolecular complex. Using these advanced methods, about 70% signals of florigen receptor are assigned.

Research Products

• [Int'l Joint Research] IOCB AS CR(チェコ)
• [Journal Article] Direct Detection of the Mercury-Nitrogen Bond in the Thymine-HgII-Thymine Base-pair with 199Hg NMR Spectroscopy (2015)
  • Author(s): Takenori Dairaku, Kyoko Furuita, Hajime Sato, Jakub Sebera, Daichi Yamanaka, Hiroyuki Otaki, Shoko Kikkawa, Yoshinori Kondo, Ritsuko Katahira, F. Matthias Bickelhaupt, Celia Fonseca Guerra, Akira Ono, Vladimir Sychrovsky, Chojiro Kojima and Yoshiyuki Tanaka
  • Journal Title: Chemical Communications Volume: 51 Issue: 40 Pages: 8488-8491
  • DOI: 10.1039/c5cc02423d
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Physicochemical Nature of Interfaces Controlling Ferredoxin NADP+ Reductase Activity through Its Interprotein Interactions with Ferredoxin (2015)
  • Author(s): Kinoshita Misaki, Kim Ju Yaen, Kume Satoshi, Sakakibara Yukiko, Sugiki Toshihiko, Kojima Chojiro, Kurisu Genji, Ikegami Takahisa, Hase Toshiharu, Kimata-Ariga Yoko, Lee Young-Ho
  • Journal Title: Biochimica et Biophysica Acta-Bioenergetics Volume: 1847 Issue: 10 Pages: 1200-1211
  • DOI: 10.1016/j.bbabio.2015.05.023
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Exploring a DNA sequence for the three-dimensional structure determination of a silver(I)-mediated C-C base pair in a DNA duplex by 1H NMR spectroscopy (2015)
  • Author(s): Takenori Dairaku, Kyoko Furuita, Hajime Sato, Yoshinori Kondo, Chojiro Kojima, Akira Ono and Yoshiyuki Tanaka
  • Journal Title: Nucleosides, Nucleotides & Nucleic Acids Volume: 34 Issue: 12 Pages: 877-900
  • DOI: 10.1080/15257770.2015.1088160
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The crystal structure of the plant small GTPase OsRacl reveals its mode of binding to NADPH oxidase. (2015)
  • Author(s): Kosami KI, Ohki I, Nagano M, Furuita F, Sugiki T, Kawano Y, Kawasaki T, Fujiwara T, Nakagawa A, Shimamoto K, Kojima C
  • Journal Title: The Journal of Biological Chemistry Volume: 289 Issue: 41 Pages: 28569-78
  • DOI: 10.1074/jbc.m114.603282
  • Description: (in press)(印刷中)
  • Peer Reviewed
• [Journal Article] Utilization of paramagnetic relaxation enhancements for high-resolution NMR structure determination of a soluble loop-rich protein with sparse NOE distance restraints (2015)
  • Author(s): Kyoko Furuita, Saori Kataoka, Toshihiko Sugiki, Yoshikazu Hattori, Naohiro Kobayashi, Takahisa Ikegami, Kazuhiro Shiozaki, Toshimichi Fujiwara and Chojiro Kojima
  • Journal Title: J. Biomol. NMR Volume: 61 Issue: 1 Pages: 55-64
  • DOI: 10.1007/s10858-014-9882-7
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ^<1>H, ^<15>N and ^<13>C resonance assignments of the conserved region in the middle domain of S. pombe Sin1 protein. (2014)
  • Author(s): Kataoka S, Furuita K Hattori Y, Kobayashi N, Ikegami T, Shiozaki K, Fujiwara T, Kojima C
  • Journal Title: Biomolecular NMR Assignments Volume: (印刷中) Issue: 1 Pages: 89-92
  • DOI: 10.1007/s12104-014-9550-6
  • Peer Reviewed
• [Journal Article] Bacterial effector modulation of host E3 ligase activity suppresses PAMP-triggered immunity in rice. (2014)
  • Author(s): Ishikawa, K., Yamaguchi, K., Sakamoto, K., Yoshimura, S., Inoue, K., Tsuge, S., Kojima, C., Kawasaki, T.
  • Journal Title: Nature Communications Volume: 5 Issue: 1 Pages: 5430-5430
  • DOI: 10.1038/ncomms6430
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Latest approaches for efficient protein production in drug discovery (2014)
  • Author(s): Toshihiko Sugiki, Toshimichi Fujiwara and Chojiro Kojima
  • Journal Title: Expert Opin. Drug Discov. Volume: 9 Issue: 10 Pages: 1189-1204
  • DOI: 10.1517/17460441.2014.941801
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] PRE and PCS techniques utilized for high-resolution protein structure determination (2015)
  • Author(s): Chojiro Kojima
  • Organizer: The 2015 International Chemical Congress of Pacific Basin Societies
  • Place of Presentation: Sheraton Waikiki (Honolulu)
  • Year and Date: 2015-12-15
  • Int'l Joint Research / Invited
• [Presentation] NMR tools developed for drug discovery (2015)
  • Author(s): Chojiro Kojima
  • Organizer: International Symposium on Structure and Folding of Disease Related Proteins
  • Place of Presentation: Seoul National University (Seoul)
  • Year and Date: 2015-12-04
  • Int'l Joint Research / Invited
• [Presentation] Utilization of Paramagnetic NMR techniques for protein structure determination (2015)
  • Author(s): Chojiro Kojima
  • Organizer: The 6th Asia-Pacific NMR Symposium
  • Place of Presentation: The Hong Kong University of Science and Technology (Hong Kong)
  • Year and Date: 2015-08-13
  • Int'l Joint Research / Invited
• [Presentation] Paramagnetic NMR techniques utilized for protein structure analysis (2015)
  • Author(s): Chojiro Kojima
  • Organizer: The 3rd Awaji International Workshop on “Electron Spin Science & Technology: Biological and Materials Science Oriented Applications”
  • Place of Presentation: The Awaji Yumebutai International Conference Center (Awaji)
  • Year and Date: 2015-06-14
  • Int'l Joint Research / Invited
• [Presentation] Drug screening using NMR to regulate protein function (2015)
  • Author(s): Chojiro Kojima
  • Organizer: The First Trilateral Workshop For Frontier Protein Studies
  • Place of Presentation: Peking University (Beijing)
  • Year and Date: 2015-04-23
  • Int'l Joint Research / Invited
• [Presentation] Paramagnetic NMR utilized for high-resolution protein structure determination (2014)
  • Author(s): Chojiro Kojima
  • Organizer: The 5th Japan-Taiwan bilateral NMR symposium
  • Place of Presentation: Hokkaido University (Sapporo)
  • Year and Date: 2014-09-29 – 2014-09-30
  • Invited