| Project/Area Number |
11671849
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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 |
Functional basic dentistry
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
TAKUMA Taishin Professor, School of Dentistry, Health Sciences University of Hokkaido, 歯学部, 教授 (40095336)
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| Co-Investigator(Kenkyū-buntansha) |
ARAKAWA Toshiya Assistant Professor, School of Dentistry, Health Sciences University of Hokkaido, 歯学部, 講師 (40306254)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥2,800,000 (Direct Cost: ¥2,800,000)
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| Keywords | SNARE hypothesis / VAMP-2 / GFP / SNAP-23 / Exocytosis / Parotid acinar cell |
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| Research Abstract |
SNARE proteins are generally accepted to be involved in the docking and fusion process of intracellular membranes. VAMP-2, syntaxin-4, and SNAP-23 are most plausible candidates of SNARE proteins for non-neuronal exocytosis. Thus, we examined the localization and intracellular trafficking of these proteins by expressing as green fluorescent protein (GFP)-tagged fusion proteins in various cells, including cultured salivary cells (HSY cells), COS-7 cells, and 3T3-L1 adipocytes. In HSY cells and COS-7 cells, VAMP-2 tagged with GFP was strongly expressed in the Golgi area. The full length of SNAP-23 with GFP was universally expressed in the cytosol except the cell nuclei. Syntaxin-4 was localized on large vesicles unidentified. When GFP-SNAP-23 and syntaxin-4 were co-expressed in these cells, SNAP-23 moved to the vesicles where GFP-syntaxin-4 was localized, suggesting that localization of SNAP-23 is determined at least partly by syntaxin-4. When these proteins were expressed in 3T3-L1 adipocytes, SNAP-23 and VAMP-2 were partly localized on the plasma membrane, although syntaxin-4 was still on the vesicles. Syntaxin-4 without transmembrane domain was not soluble but revealed as aggregates in both COS-7 cells and 3T3-L1 adipocytes. These results suggest that intracellular trafficking of SNARE proteins are regulated with other SNARE proteins or factors unidentified.
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