Anaerobic crystallization for protein crystallization

• Project/Area Number: 26650026
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: High Energy Accelerator Research Organization
• Principal Investigator: SENDA Miki 大学共同利用機関法人高エネルギー加速器研究機構, 物質構造科学研究所, 特任助教 (10707467)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: タンパク質の結晶化 / 嫌気条件下での結晶化 / 嫌気チャンバー / 結晶化の再現性向上 / タンパク質結晶化 / 結晶化スクリーニング

Outline of Final Research Achievements

It’s essential to obtain crystals with high reproducibility for an X-ray protein crystallography. Anaerobic crystallization is useful to improve reproducibility of crystallization, because oxidation of protein solutions hampered the crystal formation. We’ve established typical procedures for the anaerobic crystallization and the anaerobic crystallization was tested for several proteins. Crystals were obtained with higher reproducibility under anaerobic condition than under aerobic condition. We believe that anaerobic crystallization is effective to crystallize a various kinds of proteins.

Research Products

• [Journal Article] Improvement of Helicobacter pylori CagA crystals by the multi-step soaking method (2016)
  • Author(s): Senda, M.
  • Journal Title: Cryst. Growth Des. Volume: 16 Issue: 3 Pages: 1565-1571
  • DOI: 10.1021/acs.cgd.5b01692
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] The Lipid Kinase PI5P4Kβ Is an Intracellular GTP Sensor for Metabolism and Tumorigenesis. (2016)
  • Author(s): Sumita, K., Lo, Y., Takeuchi, K., Senda, M., Kofuji, S., Ikeda, Y., Terakawa, J., Sasaki, M., Yoshino, H., Majd, N., Zheng, Y., Kahoud, E. R., Yokota, T., Emerling, B. M., Asara, J. M., Ishida, T., Locasale, J. W., Daikoku, T., Anastasiou, D., Senda & T., Sasaki, A. T.
  • Journal Title: Molecular Cell Volume: 61 Issue: 2 Pages: 1-12
  • DOI: 10.1016/j.molcel.2015.12.011
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Crystallization and preliminary crystallographic analysis of acetophenone reductase from Geotrichum candidum NBRC 4597 (2015)
  • Author(s): Y. Sugiyama, M. Senda, T. Senda, T. Matsuda
  • Journal Title: Acta Crystallogr. Volume: F71 Issue: 3 Pages: 320-323
  • DOI: 10.1107/s2053230x15002265
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase (2014)
  • Author(s): Nishizawa A, Harada A, Senda M, Tachihara Y, Muramatsu D, Kishigami S, Mori S, Sugiyama K, Senda T, Kimura S
  • Journal Title: Biochemical Journal Volume: 462 Issue: 2 Pages: 257-265
  • DOI: 10.1042/bj20140384
  • Peer Reviewed / Open Access
• [Journal Article] Molecular mechanism of strict substrate specificity of an extradiol dioxygenase, DesB, derived from Sphingobium sp. SYK-6. (2014)
  • Author(s): K. Sugimoto, M. Senda, D. Kasai, E. Masai, M. Fukuda and T. Senda
  • Journal Title: PLoS One Volume: 9
  • Peer Reviewed / Open Access
• [Presentation] 嫌気条件下で結晶化の成功率を高める手法 (2015)
  • Author(s): 千田美紀、千田俊哉
  • Organizer: BMB2015
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2015-12-03
• [Presentation] A comprehensive strategy to obtain high quality crystals. (2015)
  • Author(s): Miki Senda, Toshiya Senda
  • Organizer: American Crystallographic Association (ACA) 2015
  • Place of Presentation: フィラデルフィア（米国）
  • Year and Date: 2015-07-29
  • Int'l Joint Research
• [Presentation] タンパク質結晶の質を効率良く改善するための結晶工学的処理の手法 (2015)
  • Author(s): 千田美紀, 千田俊哉
  • Organizer: 第15回日本蛋白質科学会年会
  • Place of Presentation: あわぎんホール（徳島県徳島市）
  • Year and Date: 2015-06-26
  • Invited
• [Presentation] フェレドキシン還元酵素BphA4の酸化還元状態依存的な構造変化と親和性調節の解明 (2014)
  • Author(s): 千田美紀
  • Organizer: 日本結晶学会平成26年度年会
  • Place of Presentation: 東京大学農学部（東京都文京区）
  • Year and Date: 2014-11-01
• [Presentation] Anaerobic crystallization for protein crystallography. (2014)
  • Author(s): 千田美紀
  • Organizer: 23rd Congress of the IUCr
  • Place of Presentation: Palais des Congres de Montreal (カナダ、モントリオール）
  • Year and Date: 2014-08-11
• [Presentation] 嫌気条件下でのタンパク質結晶化 (2014)
  • Author(s): 千田美紀
  • Organizer: 第14回日本蛋白質科学会
  • Place of Presentation: ワークピア横浜（神奈川県横浜市）
  • Year and Date: 2014-06-26