| Project/Area Number |
13460152
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | RIKEN (The Institute of Physical and Chemical Research) |
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Principal Investigator |
MATSUOKA Ken Laboratory for Structural Construction Laboratory Head, 形態構築研究チーム, チームリーダー (40222294)
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| Co-Investigator(Kenkyū-buntansha) |
YUASA Koji Laboratory for Structural Construction Research Scientist, 形態構築研究チーム, 研究員 (60342870)
TAKEUCHI Masaki Molecular Membrane Biology Laboratory Contract Researcher, 生体膜研究室, 協力研究員 (00332271)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥15,500,000 (Direct Cost: ¥15,500,000)
Fiscal Year 2002: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2001: ¥11,000,000 (Direct Cost: ¥11,000,000)
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| Keywords | Protein transport / Protein modification / Reconstitution / Plant / Vacuole / Membrane protein / Solubilization / Transport vesicle / タバコ / 分泌系 / EST / 再構成系 / 踏査付加 / ヒドロキシプロリン / 表面構造 |
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| Research Abstract |
Genetically modified crops are expected as factories to produce useful proteins of animal origin because of the lower cost of production than animals and microorganisms. Most of the useful proteins from animals are secretory proteins and these proteins are modified with glycans in the Golgi appratus after synthesis at the ER, and then trasnported to the vacuole/lysosome or extracellular space. Molecular mechanism of protein sorting and protein modification in the Golgi apparatus are different in organisms belonging to different kingdom ; thus, production of these animal-oriented proteins in plants as active forms require the control of plant secretory pathway by suppressiing the plant sorting and/or modification mechanism. In this research, we conducted the analysis of protein sorting apparatus from yeasts as well as the analysis of protein sorting in plants by developing in vitro system to generate transport vesicles from isolated membranes and cytosol. We also carried out analyses on glycosylation and proline hydroxylation of secretory proteins in plants. For these purposes we aimed to develop a novel methods to analyze membrane proteins. Based on these works we got the following results : 1) Definition of the topological orientation of coat proteins of COPII vesicles. 2) Role of a GTP-ase Arf1p on the protein transport in plants. 3) Characterization of proryl hydroxylase from plants. In addition we developed a rapid system to determine the solubilization condition of membrane proteins.
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