Identification of intracellular molecular interactions under living conditions
| Project/Area Number |
15K16562
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Chemical biology
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| Research Institution | Kochi University |
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Principal Investigator |
YAMAGUCHI Arisa 高知大学, 医学部, 日本学術振興会 特別研究員 (90553157)
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| Project Period (FY) |
2015-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | プロテオミクス / 分子アッセンブリー |
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| Outline of Final Research Achievements |
We investigated the issue of whether the EMARS system with fluorescein tyramide (FT) is applicable to analysis of co-clustered molecules in the intracellular organelles. To this end, we constructed HRP or APEX targeted to the ER, Golgi and secretory vesicle. The expressed HRP or APEX in the intracellular organelles were co-localized with fluorescein labeling resulting from the EMARS reaction. These observations indicate that the EMARS reaction with FT as the labeling reagent is applicable to the analysis of co-clustered molecules in the intracellular organelles.
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| Academic Significance and Societal Importance of the Research Achievements |
「HRP発現型・高特異性EMARS法」は、特異性・反応効率性・感度が確保されており、対象分子が微量でも検出可能である。よって、従来、細胞のホモジェネートやオルガネラの分画時に損失・離散していた微量な機能性分子を同定できる可能性が高い。また、従来ホモジェネート時に膜に張り付き、誤同定されていたであろう「コンタミ分子」が検出されない。本研究で確立された細胞内EMARS法を用いれば、多種多様な細胞の細胞内分子機構の解明に貢献するとともに、その解明を目指す生命科学分野の研究者に新しい展望を与えるものと期待される。
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Report
(6 results)
Research Products
(5 results)
