| Project/Area Number |
22657029
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Structural biochemistry
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
HIROSE Shigehisa 東京工業大学, 大学院・生命理工学研究科, 教授 (10134199)
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| Co-Investigator(Renkei-kenkyūsha) |
KATO Akira 東京工業大学, 大学院・生命理工学研究科, 助教 (40311336)
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| Project Period (FY) |
2010 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥3,360,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2010: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | 脂質 / タンパク質 / 糖鎖 / 細胞膜 / 脂質ラフト / ガングリオシド / GOTO細胞 / モノクローナル抗体 / フロチリン / エンドサイトーシス / 神経芽腫 / シクロデキストリン / GM2 |
|---|
| Research Abstract |
We raised a monoclonal antibody which enables the visualization of ganglioside GM2-rich microdomains as highly punctate dots on a human neuroblastoma cell, GOTO. Using this antibody, we showed that raft-like microdomains of GM2(GM2rafts) were colocalized with flotillin-2/reggie-1-positive microdomains. Prelabeling of GM2 rafts with a fluorescent probe, Alexa555. Anti-GM2, demonstrated internalization and delivery of the raft. flotillin assembly to the recycling endosomes. Depletion of cholesterol with methyl-β-cyclodextrin(MβCD) resulted in disruption and targeting of GM2 rafts to lysosomes. These results suggest that in GOTO cells, GM2 rafts are actually present in association with flotillin and undergo internalization into distinct pools of endosomal vesicles depending on their cholesterol contents.
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