| Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
|
|---|
| Outline of Final Research Achievements |
Thymidine phosphorylase (TP) is a unique metabolic control enzyme, TP can be used to change the nucleobase moiety of thymidine to a modified uracil substituted at the C5 position. The cross-coupling reaction by palladium catalyst such as Sonogashira coupling is convenient tool for introducing a fluorescent group at C5 position of uracil. We reasoned that TP and Pd catalyzed cross-coupling reaction could be used to easily obtain many different nucleosides containing a fluorescent group. We attempted to perform the base-exchange reaction by a hydrophobic modified uracil in buffer containing a polar organic solvent such as DMSO with TP. The reaction conversion was drastically enhanced as the concentration of DMSO increased up to 40 % (57 % conversion for U4C compound). Docking simulations using MF myPresto showed that uracil moiety of the substrate bound to the active site of TP, the fluorescent moiety linked to the C5 position of the uracil located outside the surface of the enzyme.
|
