2020 Fiscal Year Annual Research Report
Site-Selective Protein Chemical Modification of Exposed Tyrosine Residues Using Tyrosine Click Reaction
Publicly Offered Research
| Project Area | Non-equilibrium-state molecular movies and their applications |
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| Project/Area Number |
20H05438
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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) |
2020-04-01 – 2022-03-31
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| Keywords | CO release reaction / Initial intermediate / TR-SFX method |
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| Outline of Annual Research Achievements |
The proposed research is progressing well and the current results are described in two parts as follows.(1) Capturing initial intermediates of a CO release reaction within a protein crystal: We Immobilized Mn(CO)3 as a light-induced CO release reaction center inside the Lysozyme microcrystals and studied the light reactions by TR-SFX with varying time and light dose. We found that Mn(CO)3 form an octahedral geometry with two additional water molecules. At the initial stage of reaction, the axial CO released first and we successfully captured the Mn(CO)2 intermediate species. With increasing delay time up to 1ms, we observed the complete release of Mn(CO) unit from the protein. During such chemical reaction, we determined the conformational changes in the surrounding Arg14 residue. Overall, the work is about to finish and we are applying this methodology to new reaction systems. such as gold nanocluster formation etc.(2) Observing gold nanocluster formation within the symmetric channel of a ferritin cage: In this research, we are studying to explore the mechanism of gold cluster formation in protein cavity. At first, we immobilized the Au atoms in the ferritin cage and determined thepositions of Au atoms in the protein structure. Then, we optimized the photoreduction of Au atoms to see movements of Au atoms along with side chain conformational changes during the formation of the nanocluster by usual X-ray crystallography. In the next plan, we will apply TR-SFX method to study the intermediates in short intervals to understand the detail mechanism.
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| Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
The proposed research is progressing well more than we expected. We have established the proposed concept and currently applying to several reaction systems.
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| Strategy for Future Research Activity |
Our future research plan related to the proposed research is as follows.
(1) Submission of the current research result on exploring CO release mechanism by TR-SFX.
(2) We are extending our proposed concept to other reaction systems such as gold nanocluster formation mechanism in the symmetric interfaces of a ferritin protein cage. We have established the initial optimization of the reaction and structure determination by X-ray crystallography. In our next plan, we will apply this system to TR-SFX to explore the detailed mechanism of cluster formation.
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