Investigation of molecular mechanism of transphosphorylation by a phosphatidylinositol kinase

• Project/Area Number: 15H06855
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: High Energy Accelerator Research Organization
• Principal Investigator: SATO Tomomi 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, 博士研究員 (20759211)
• Research Collaborator: TAKEUCHI Ko 産業技術総合研究所, 創薬分子プロファイリングセンター, 主任研究員 ; SASAKI Atsuo University of Cincinnati, Associate Professor
• Project Period (FY): 2015-08-28 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 構造生物学 / キナーゼ / 相互作用解析 / イノシトールリン脂質 / 結晶化 / PI5P4K / リン酸化反応機構 / X線結晶構造解析

Outline of Final Research Achievements

In this research project, I performed following three experiments in order to elucidate catalytic mechanism of phosphorylation by an inositol phospholipid kinase, PI5P4K beta. 1) Interaction analysis between PI5P4K beta and lipid/nucleotide by SPR, 2) X-ray crystallographic structural analysis in complex with inhibitors, 3) Crystallization of PI5P4K alpha/gamma. From the results of the SPR analysis, I elucidated that the interaction between background lipid and PI5P4K beta is relatively strong (micro molar order) while the intersection between substrate (PI5P) and PI5P4K beta is predicted to be weak. I also succeeded in observation of electron density of 3 inhibitors and crystallization of PI5P4K alpha.

Research Products

• [Int'l Joint Research] シンシナティー大学(米国)