Comprehensive analyses of protein interactors of disease associated prion protein
| Project/Area Number |
16K08038
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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 |
Veterinary medical science
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| Research Institution | National Agriculture and Food Research Organization |
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Principal Investigator |
Iwamaru Yoshifumi 国立研究開発法人農業・食品産業技術総合研究機構, 動物衛生研究部門, ユニット長 (20355142)
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| Research Collaborator |
Nakanishi Jun
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | プリオン / 異常プリオン蛋白質 / 培養細胞 / EMARS / 近接標識 / 細胞膜 / FITC / GT1-7細胞 / 蛋白質 / 標識 / 感染細胞 / 異常型プリオン蛋白質 / 細胞表面 / 正常プリオン蛋白質 / EMARS 反応 / 共局在 / 細胞 / 脂質ラフト / プリオン病 / プリオン持続感染細胞 / 神経細胞 |
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| Outline of Final Research Achievements |
To identify the candidate molecules capable of interacting with abnormal prion protein (PrPSc) and facilitating PrPSc formation at the cell membrane, we applied a biochemical labelling method termed ‘enzyme-mediated activation of radical sources (EMARS). First, the optimal conditions to label cell-surface PrPSc in viable cells were investigated. Using an approach based on a recent study describing amino-proximal anti-PrP antibodies react natively with cell surface PrPSc in its physiological context, we evaluated several amino-proximal anti-PrP antibodies and optimized the immunocytochemical conditions for immunolabeling aggregated PrPSc on the cell-surface of prion-infected GT1-7 cells. EMARS was applied to the prion-infected GT1-7 cells whose cell surface PrPSc was pre-labelled with one of the amino-proximal anti-PrP antibodies.Co-immunoprecipitation and Western blot analysis showed several proteins were labelled with FITC in a PrPSc-specific manner.
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| Academic Significance and Societal Importance of the Research Achievements |
生理的条件下で、プリオン感染細胞膜上の異常プリオン蛋白質を効率的に標識可能となった。この免疫細胞化学法とEMARS法を組み合わせ、生細胞上の異常プリオン蛋白質に近接する分子群を特異的に標識可能であること示した。異常プリオン蛋白質と共局在する分子群を同定し、治療標的とすることで、未だ有効な治療法のないプリオン病に対する予防、治療法の開発につながる可能性がある。
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Report
(4 results)
Research Products
(2 results)
