NMR analyses of the structural equilibrium in tyrosine kinases

• Project/Area Number: 26840014
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Keio University (2015); Hokkaido University (2014)
• Principal Investigator: YOKOGAWA MARIKO 慶應義塾大学, 薬学部, 助教 (60648020)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: キナーゼ / NMR / チロシンキナーゼ

Outline of Final Research Achievements

Tyrosine kinases are major targets for anticancer drug. To understand the dynamic regulation of tyrosine kinases, NMR analyses of the structural equilibrium were performed. In this study, E coli. expression system and purification protocol for c-Src, Fyn, and c-Abl were successfully established. NMR spectral changes upon phosphorylation of c-Src and upon binding to drugs were observed. NMR approach would be useful for understanding the dynamic processes in the regulation of tyrosine kinases.

Research Products

• [Journal Article] Nuclear magnetic resonance analysis of the conformational state of cancer mutant of fibroblast growth factor receptor 1 tyrosine kinase domain. (2016)
  • Author(s): Kobashigawa Y., Amano S., Yoza K., Himeno R., Amemiya S., Morioka H., Yokogawa M., Kumeta H., Schlessinger J., and Inagaki F.
  • Journal Title: Genes to Cells Volume: 21(4) Issue: 4 Pages: 350-357
  • DOI: 10.1111/gtc.12345
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structural basis for the regulation of enzymatic activity of Regnase-1 by domain-domain interactions. (2016)
  • Author(s): Yokogawa M., Tsushima T., Noda NN., Kumeta H., Enokizono Y., Yamashita K., Standley DM., Takeuchi O., Akira S., and Inagaki F.
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 22324-22324
  • DOI: 10.1038/srep22324
  • NAID: 120005818281
  • Peer Reviewed / Open Access
• [Journal Article] NMR Method for Characterizing Microsecond-to-Millisecond Chemical Exchanges Utilizing Differential Multiple-Quantum Relaxation in High Molecular Weight Proteins. (2016)
  • Author(s): Toyama Y., Osawa M., Yokogawa M., and Shimada I.
  • Journal Title: J. Am. Chem. Soc. Volume: 138(7) Issue: 7 Pages: 2302-2311
  • DOI: 10.1021/jacs.5b12954
  • NAID: 40021421900
  • Peer Reviewed
• [Journal Article] Kobashigawa Y, Amano S, Yokogawa M, Kumeta H, Morioka H, Inouye M, Schlessinger J, Inagaki F. (2015)
  • Author(s): Structural analysis of the mechanism of phosphorylation of a critical autoregulatory tyrosine residue in FGFR1 kinase domain.
  • Journal Title: Genes Cells. Volume: 20 Issue: 10 Pages: 860-870
  • DOI: 10.1111/gtc.12277
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] ポリA上でのPABP多量体化の構造生物学的解析 (2016)
  • Author(s): 沢崎綾一, 今井駿輔, 横川真梨子, 臼井遥子, 星野真一, 大澤匡範
  • Organizer: 日本薬学会第136回年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-03-26
• [Presentation] Gタンパク質三量体によるGタンパク質共役型内向き整流性カリウムチャネル活性制御機構の構造生物学的解明 (2015)
  • Author(s): 加納花穂, 外山侑樹, 岩橋優太, 間瀬瑶子, 横川真梨子, 大澤 匡範, 嶋田一夫
  • Organizer: 第38回日本分子生物学会年会 第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] 多量子遷移の緩和速度の差を利用した高分子量タンパク質のマイクロ秒からミリ 秒オーダーの化学交換の解析法の開発 (2015)
  • Author(s): 外山侑樹, 大澤匡範, 横川真梨子, 嶋田一夫
  • Organizer: 第54回NMR討論会
  • Place of Presentation: 千葉工業大学（千葉県習志野市）
  • Year and Date: 2015-11-06
• [Presentation] Regnase-1によるmRNA切断機構の構造生物学的解析 (2014)
  • Author(s): 横川 真梨子、津嶋 崇、野田 展生、久米田 博之、足立 わかな、榎園 能章、山下 和男、Daron M. Standley、竹内 理、審良 静男、稲垣 冬彦
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Gタンパク質三量体によるGタンパク質共役型内向き整流性カリウムチャネル活性制御機構の構造生物学的解明 (2014)
  • Author(s): 加納 花穂, 外山 侑樹, 岩橋 優太, 間瀬 瑶子, 横川 真梨子, 大澤 匡範, 嶋田 一夫
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Regnase-1によるmRNA切断機構の構造生物学的解析 (2014)
  • Author(s): 横川 真梨子、津嶋 崇、野田 展生、久米田 博之、足立 わかな、榎園 能章、山下 和男、Daron M. Standley、竹内 理、審良 静男、稲垣 冬彦
  • Organizer: 第53回NMR討論会
  • Place of Presentation: 大阪大学（大阪府吹田市）
  • Year and Date: 2014-11-04 – 2014-11-06
• [Presentation] Structural Basis for Regulation of G Protein-activated Inwardly Rectifying Potassium Channel 1 (GIRK1) by G Proteins (2014)
  • Author(s): Osawa M, Mase Y, Yokogawa M,Takeuchi K, Shimada I
  • Organizer: 第52回日本生生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Structural Basis for Regulation of G Protein-activated Inwardly Rectifying Potassium Channel 1 (GIRK1) by G Proteins. (2014)
  • Author(s): Osawa M, Mase Y, Yokogawa M,Takeuchi K, Shimada I
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Dallas （USA）
  • Year and Date: 2014-08-24 – 2014-08-29
• [Presentation] Structural analyses of the gating mechanism of inwardly rectifying potassium channel. (2014)
  • Author(s): Toyama Y, Osawa M, Yokogawa M, Shimada I
  • Organizer: International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: Dallas （USA）
  • Year and Date: 2014-08-24 – 2014-08-29