| Project/Area Number |
06453193
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
医薬分子機能学
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| Research Institution | TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY |
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Principal Investigator |
HATANAKA Yasumaru Research Inst. Wakan-Yaku, TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY Associate Professor, 和漢薬研究所, 助教授 (30111181)
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| Co-Investigator(Kenkyū-buntansha) |
TEZUKA Yasuhiro Research Inst. Wakan-Yaku, TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY Assistan, 和漢薬研究所, 助手 (70236975)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1995: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1994: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | photoaffinity labeling / opioid receptor / diazirine / carbene / chemiluminescence / avidin / biotin / structure-function relation ship / アジド / 微生物代謝成分 |
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| Research Abstract |
Research Purpose : Recent success in the construction of chimeric opioid receptors demonstrated the presence of several extra cellular regions which could be responsible for the distinct ligand-binding profiles of the each type of opioid receptors. However, it is generally impossible to exclude potential conformational changes which result in alterations of the ligand binding. The purpose of this research is the development and application of diazirine based photoprobes for the analysis of opioid binding sites.
Results : A convenient synthesis of m- and p-CF_3-diazirinylbenzoic acid was developed. Using these photoreactive moieties, a novel pair of photoprobes which only differ in the carbene generating sites were synthesized. This structural feature of the probes are potentially useful to map the different sites within the ligand binding pocket. For avoiding the use of radioactive probes, a novel biotinylated photoreactive naltrexone analog was also developed. All new probes bearing diazirine bind reversibly with high affinity at mu-, delta-, and kappa- receptors. The photoaffinity labeling of recombinant delta-receptor protein was investigated with the biotinyl probe. The visualization of target molecule was successfully accomplished by the chemiluminescent detection of photochemically biotinylated proteins.
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