| Project/Area Number |
09680705
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Kansai Medical University |
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Principal Investigator |
YAMAMOTO Akitsugu Kansai Medical University, Department of Physiology, lecture, 医学部, 講師 (30174775)
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| Co-Investigator(Kenkyū-buntansha) |
MASAKI Ryuichi Kansai Medical University, Department of Physiology, lecture, 医学部, 講師 (70140283)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | C-terminal-anchored membrane protein / endoplasmic reticulum / Golgi apparatus / microsomal aldehyde dehydrogenase / HPC-1 / syntaxin 1a / green fluorescent protein / plasma membrane / vesicular traffic / GFP / 局在化 / 膜蛋白質 / 滑面小胞体 / 肝臓 / 小腸 / 選別 |
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| Research Abstract |
Carboxyl (C)-terminal-anchored membrane proteins are anchored to the membrane by a hydrophobic domain at its C-terminus, and play many important roles distributing on many kinds of organella such as the endoplasmic reticulum (ER), Golgi apparatus, the plasma membrane, endosomes, synaptic vesicles, mitochondria and so on. For instances, microsomal aldehyde dehydrogenase (msALDH) localizes on the ER membrane as a enzymes, while HPC-1(syntaxin 1a) localizes on the plasma membrane as an important vesicular traffic protein. In this research project, we carried out following studies in order to elucidate mechanism of the localization of the C-terminal anchored membrane proteins.
1. Mechanism of the localization of HPC-1 on the plasma membrane. We investigated mechanism of localization of HPC-1 on the plasma membrane by transfecting cDNA for this protein into COS cells. As the results, it was shown that HPC-1 is integrated into ER membrane first, and then transported to the plasma membrane through Golgi apparatus. By transfecting chimeric protein with maltose binding protein (MBP) and C-terminal hydrophobic domain of HPC-1 or msALDH, it was suggested that localization signal for the plasma membrane or the ER exists on the C-terminal hydrophobic domain of HPC-1 or msALDH, respectively.
2. Retention mechanism of msALDH on the ER. A Chimeric green fluorescent protein (GFP) containing the C-terminal hydrophobic domain of msALDH (GFP/msALDH) was localized on the ER when it was expressed in COS cells or NRK cells. ER localization of GFP/msALDH was not changed by AlFィイD24ィエD2ィイD1-ィエD1treatment which inhibits recycling between ER and Golgi apparatus. This suggests that GFP/msALDH is retained on the ER by static retention mechanism.
3. Distribution of msALDH in rat intestine. We found that msALDH are localized on the smooth ER of the adsorptive cells of rat small intestine at high density.
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