2012 Fiscal Year Final Research Report
Development of high-resolution pulse-chase method by use of click chemical reaction
| Project/Area Number |
23657126
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Cell biology
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| Research Institution | Nagoya University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Michitaka 名古屋大学, 医学系研究科, 助教 (80456649)
OZATO Nami 名古屋大学, 医学系研究科, COE 特任助教 (60547454)
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| Project Period (FY) |
2011 – 2012
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| Keywords | 膜脂質 / 凍結割断レプリカ標識法 / クリック反応 |
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| Research Abstract |
Cells cultured with precursor molecules bearing an alkyl or an azido group were quick-frozen and freeze-fracture replicas were subjected to click chemical reaction to label membrane lipids and proteins for electron microscopy. The labeling for phosphatidylcholine increased in a time-dependent manner after the precursor treatment. In budding yeast, the labeling density in the endoplasmic reticulum, nuclear membrane, mitochondrion, and vacuole was equivalent in the two membrane leaflets, whereas that in the plasma membrane showed clear asymmetry.
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