2014 Fiscal Year Annual Research Report
Ｋｅａｐ１－Ｎｒｆ２の相互作用を阻害する擬天然物特殊ペプチドのプロドラッグ化
| Project/Area Number |
13F03730
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| Research Institution | The University of Tokyo |
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Principal Investigator |
菅 裕明 東京大学, 理学(系)研究科(研究院), 教授 (00361668)
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| Co-Investigator(Kenkyū-buntansha) |
GEIERMANN Anna-Skrollan 東京大学, 理学(系)研究科(研究院), 外国人特別研究員
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | 遺伝記号 / 特殊ペプチド / tRNAアルシ化 / 翻訳 |
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| Outline of Annual Research Achievements |
I learned new techniques and I am now confident with the methods used for selection technology. I completed one selection against the chemokine receptor CXCR7. 12 macrocyclic peptides were identified and synthesized and are now being used in co-crystallization experiments. A new selection project against the mitochondrial membrane transporters ATOM of trypanosomes and human TOM40 was started. Major problems were aggregation of ATOM and lack of enrichment of positive binders. Finally enrichment was observed and sequencing is performed now. Furthermore, 6 MS2-tagged ribosomes containing different mutations were successfully produced by site-directed mutagenesis.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The CXCR7 project was successfully finished from my side. I selected 12 peptides using the RaPID system and synthesized these peptides. Currently they are used for crystallization experiments by the collaborator. The current selection project against the protein TOM40 is going very well after overcoming initial problems and recent sequencing results gave high convergence. Here, the selected peptide sequences will be synthesized soon and sent to the collaborator. The second selection project against Atom started with problems of protein aggregation. However, together with the collaborators I am now working on overcoming these problems in order to finish this project successfully.
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| Strategy for Future Research Activity |
I plan to finish the selection against proteins ATOM and TOM40. Standard sequencing as well as next-generation sequencing will be performed. If the sequencing data is good, I will synthesize and purify the peptides and they can be sent to the collaborators.
I plan to continue working on the ribosome project. I will purify the mutant ribosomes, do translation assays and check the incorporation of D-amino acids to study the mutant ribosomes ability to incorporate not only single but also several consecutive D-amino acids. This project could help to evaluate the role of the mutated nucleotides in D-amino acid incorporation.
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