| Project Area | Non-equilibrium-state molecular movies and their applications |
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| Project/Area Number |
22H04744
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| Research Category |
Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
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| Allocation Type | Single-year Grants |
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| Review Section |
Complex systems
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
Maity Basudev 東京工業大学, 生命理工学院, 特任助教 (60815421)
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| Project Period (FY) |
2022-04-01 – 2024-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥11,180,000 (Direct Cost: ¥8,600,000、Indirect Cost: ¥2,580,000)
Fiscal Year 2023: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2022: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
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| Keywords | Ferritin cage / Copper complex / ferritin / Metal complex / Light reaction / Ferritin cage crystal / Cu(I) complex / Structural distortion / Serial crystallography |
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| Outline of Research at the Start |
Excited state structural dynamics of transition metal complexes are important not only for applications but also for rethinking the fundamental concepts. Ultrafast spectroscopy is the common method for such studies. However, real-space determination of the structural changes in the excited state remained challenging due to transient nature of the intermediate(S). In this proposal, we aim to use the TR-SFX method for observing structural dynamics of a tetrahedral Cu(I)-phen complex in ultrafast timescale.
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| Outline of Annual Research Achievements |
Under this project, we have the following achievements.
We prepared the ferritin mutants with a designed cavity at the 2 fold symmetric interface and immobilized the Cu-phenanthroline complex. The composite was crystallized and determined the X-ray structure. In an additional work, we achieved to study the reaction dynamics of a MnCO3 complex in RNaseA protein and determined the XFEL structure and plan to study the dynamics.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
Our previously designed mutant showed copper complex binding as evident from quantitative analysis and red colored crystals. However, we did not observe enough electron density in the X-ray structure. Observing good electron density is essential to study structural dynamics by TR-SFX. Therefore, we are redesigning the protein cavity for strong binding of metal complex and expected to get good electron density. During this time, we also developed an additional MnCO3 complex RNaseA protein crystal to show the protein side-chain dynamics with changes in the coordination dynamics.
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| Strategy for Future Research Activity |
These are the plan for our research scheme.
Preparation of newly designed mutants which contains a cavity to accommodate copper(I)-phenanthroline complex. Crystallize the ferritin composite and screen the X-ray structure for use in XFEL. We also plan to prepare phenanthroline derivatives to fit into the designed cavity to study the dynamics of the metal complex.
In the second work, we are going to use the microcrystals of RNaseA-MnCO3 complex for SACLA measurement to explore the protein side chain dynamics during the reaction. We already optimized the microcrystallization of RNaseA-MnCO3 and determined the XFEL structure which showed all the coordination of the metal complex.
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