| 研究領域 | 高速分子動画法によるタンパク質非平衡状態構造解析と分子制御への応用 |
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| 研究課題/領域番号 |
22H04744
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| 研究機関 | 東京工業大学 |
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研究代表者 |
Maity Basudev 東京工業大学, 生命理工学院, 特任助教 (60815421)
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| 研究期間 (年度) |
2022-04-01 – 2024-03-31
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| キーワード | ferritin / Metal complex / Light reaction |
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| 研究実績の概要 |
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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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
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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| 今後の研究の推進方策 |
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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