| Project/Area Number |
60470143
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemical pharmacy
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| Research Institution | Hokkaido University |
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Principal Investigator |
KANAOKA Yuichi Fac. of Pharmaceutical Sciences, Hokkaido University, 薬学部, 教授 (50001029)
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| Co-Investigator(Kenkyū-buntansha) |
HATANAKA Yasumaru 北海道大学, 薬学部, 教務職員 (30111181)
SATO Eisuke Fac. of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助手 (10001059)
TANIZAWA Kazutaka Fac. of Pharmaceutical Sciences, Hokkaido University, 薬学部, 助教授 (90001049)
NAKAYAMA Hitoshi Fac, of Pharmaceutical Sciences, Hokkaido University (Yoshihara,Te), 薬学部, 助手 (70088863)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1987: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1986: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1985: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | Sodium channel / Biospecific Reagents / Tetrodotoxin / Photoaffinity Labeling / 新しい光反応性基 / テトロドトキシン誘導体 / ナトリゥムチャンネル / 電気ウナギ / 結合実験 / 特異的ラベル / 250Kタンパク |
|---|
| Research Abstract |
As an approach to reveal the molecular architecture of the functional components in the sodium channel, we applied the photoaffinity labeling technique using photoactivable tetrodotoxin (TTX) derivatives. One of the TTX derivatives, Nap-en-TTX 1, apparently labeled the eel sodium channel of 250 kDa polypeptide. However, during the separation and isolation steps of the labeled peptides, there appeared two proplems which prevent from identifying the labeled sites. One of the problems is their lable labeling and the other is the wide-spread of the labeling.
So we synthesized the TTX derivatives containing other types of the photosensitive moieties such as a phenyldiazirine and a fluoro-nitophenoxy group. All the compound synthesized show comparable affinities to tetrodotoxin itself in binding to the sodium channel prepration. But the extents of photoincorporation and their stabilities during purification are markedly different between the compounds, Junged from several points of these criteria we have chosen the compound 2 containing a phenyldiazirine is most suitable one, and the further study using 2 is currently under the way.
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