| Project/Area Number |
25350984
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Chemical biology
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
Kimura Tadashi 国立研究開発法人産業技術総合研究所, 創薬基盤研究部門, 主任研究員 (60344214)
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| Co-Investigator(Kenkyū-buntansha) |
Kubo Tai 国立研究開発法人産業技術総合研究所, 創薬分子プロファイリング研究センター, 副センター長 (10178030)
Kameyama Kimihiko 国立研究開発法人産業技術総合研究所, 企画本部, 総括企画主幹 (50224697)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | ペプチド / カルシウムチャネル / PERISS法 / 電気生理 / 大腸菌 / ICK / イオンチャネル / 電位センサー / 進化分子工学 / パッチクランプ / パドルキメラ |
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| Outline of Final Research Achievements |
We expressed a eukaryotic voltage-gated ion channel Kv2.1 in Escherichia coli using prokaryotic codon, and prepared giant spheroplasts large enough for the patch clamp. Human Kv2.1 currents were successfully recorded from giant spheroplasts with the automated system. ProTx-I, ProTx-II, GTx1-15, and GsMTx-4 were spider-derived ICKs and incubated with pepsin, trypsin, chymotrypsin, and elastase in physiological conditions to find that all tested peptides were resistant to pepsin, and ProTx-II, GsMTx-4, and GTx1-15 showed resistance to all tested proteases. These results become the basic knowledge of this theme. After construction of paddle chimeras, we have performed multiple peptide screening by PERISS method and we have obtained several results.
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