2016 Fiscal Year Annual Research Report
| Project/Area Number |
15F15333
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| Research Institution | The University of Tokyo |
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Principal Investigator |
菅 裕明 東京大学, 大学院理学系研究科(理学部), 教授 (00361668)
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| Co-Investigator(Kenkyū-buntansha) |
OBEXER RICHARD 東京大学, 大学院理学系研究科(理学部), 外国人特別研究員
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| Project Period (FY) |
2015-11-09 – 2018-03-31
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| Keywords | ペプチド |
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| Outline of Annual Research Achievements |
Reactive peptides against the SNAP and Halo tag that were discovered in the previous year through the RaPID platform were further analyzed and their reactivity was confirmed. Intriguingly, these peptides showed cross reactivity for both probes, indicating that these peptides are strong nucleophiles. In collaboration with the lab of Prof. Tamp, I performed selections against the ABC-transporters ‘human TAP’ and ‘bacterial Tmr’. The ultimate goal of this project is to find peptides that act as stabilizing ligands for crystallization. In case of Tmr, selections were performed against the inward and outward facing conformations leading to cyclic peptides that can distinguish both conformations. The peptides were synthesized, purified and have been handed over to the collaborator. The peptides are current evaluated by the collaborator group and attempts for the crystallization is under way.
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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 reactive peptides have been synthesized and will be analyzed in depth. Since previous selection results were already reconfirmed, characterization will reveal the biotechnological scope of these peptides, including the potential for preparing biohybrid materials and catalysts. The results achieved for the TAP transporter are highly encouraging, especially the fact that the selected peptides can distinguish the two confirmations of the protein. Additional evolution of these peptides should further promote their potential to differentiate between the two conformations. To this end, we considered the project is moving forward as expected.
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| Strategy for Future Research Activity |
The reactive peptides will be analyzed with a focus on kinetics, side reactions and substrate scope. For this purpose I will perform extensive analytical HPLC and MS studies. In addition, I will fuse the sequence to a variety of proteins (C- or N-terminally). These results will indicate the extent to which the peptides will be suitable for protein labeling and immobilization. In addition, I will test whether these peptides can be used to modify solid supports or plastics such as PVC. Due to their highly nucleophilic character I will test them as catalysts for e.g. the Rauhut-Currier reaction or Michael addition. The peptides that were raised against the ABC transporters will be further matured to increase their ability to distinguish the inward and outward facing confirmations.
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Research Products
(2 results)
