| Project/Area Number |
10215206
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (B)
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Nagahama Institute of Bio-Science and Technology (2001)
Kansai Medical University (1998-2000) |
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Principal Investigator |
YAMAMOTO Akitsugu Department of Bio-Science, Professor, バイオサイエンス学部, 教授 (30174775)
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| Project Period (FY) |
1998 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥32,500,000 (Direct Cost: ¥32,500,000)
Fiscal Year 2001: ¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 2000: ¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1999: ¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 1998: ¥7,500,000 (Direct Cost: ¥7,500,000)
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| Keywords | endoplasmic rericulum / Golgi appatatus / transport vesicle / immuno-electron microscopy / microsmal aldehyde dehydrogenase / synaptotagmin / crystalloid endoplasmic reticulum / vesicular transport / syntaxin / HPC-1 / COS細胞 / VSV-G蛋白質 / 膜蛋白質 / ミクロゾーム型アルデヒド脱水素酵素 |
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| Research Abstract |
Microsomal aldehyde dehydrogenase (msALDH) is a C-terminal anchored membrane protein, and anchors to the membrane of endoplasmic reticulum (ER) by a hydrophobic region at the C-terminus. We reported that the C-terminal 35 amino acids possess the signal for targeting of msALDH to the ER. In the present study, a chimera protein in which the C-terminal 35 amino acids of msALDH was added to the C-terminus of green fluorescence protein (GFP) was expressed in COS cells and NRK cells, and intracellular distribution of the chimera protein was examined by highly sensitive immuno-electron microscopy. The chimera protein was densely localized in the ER and nuclear membrane, but was not detected in the transport vesicles between the ER and the Golgi apparatus as well as the Golgi apparatus. This fact suggests that msALDH is excluded from transport vesicles when they form from the ER, and statically retained in the ER.
We have reported that overexpression of msALDH results in the formation of the crystalloid ER in cultured cells. In this study, we found that crystalloid ER forms by expressing a mutant of synaptotagmin (Syt), a family of type I membrane proteins. Wild Syt IV was localized in the Golgi apparatus. Deletion of C-terminal 17 amino acids of Syt 4 prevented die Golgi localization and induced crystalloid ER, possibly oligomerization of malfoled protein in the ER membrane.
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