| Project/Area Number |
13670275
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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 |
Bacteriology (including Mycology)
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| Research Institution | Ehime University |
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Principal Investigator |
SHINOMIYA Hiroto Ehime University School of Medicine, Dept. of Immunology and Host Defenses, Associate Professor, 医学部, 助教授 (80162618)
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| Co-Investigator(Kenkyū-buntansha) |
ASANO Yoshihiro Ehime University School of Medicine, Dept. of Immunology and Host Defenses, Professor, 医学部, 教授 (70114353)
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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 |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | p65 / L-plastin / phosphorylation / host defense / cytoskeleton / cell adhesion / actin / recombinant protein / immunostaining / P65 |
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| Research Abstract |
We previously identified a 65-kDa protein (p65) that was phosphorylated in activated macrophages. It has turned out to be a murine homologue of human L-plastin that was identified as a novel protein in human cancer cells. P65/L-plastin is characterized by a series of Ca^<2+>-, calmodulin- and actin-binding domains, and is thought to play a crucial role in leukocytes and cancer cells. We have expressed a recombinant γ p65/L-plastin in E. coli that binds to b-actin and prepared high-titer antibodies using large amounts of the protein as immunogen. Anti-rp65/L-plastin antibodies recognize native p65/L-plastin as well as rp65/L-plastin and have enabled us to detect fine structures of intracellular p65/L-plastin, and it was found that its localization was extensively changed by stimulation with bacterial components. We further developed an enzyme-linked immunosorbent assay system and a flow cytometry method using these reagents, which made it possible to measure antibodies including autoantibodies against p65/L-plastin and to evaluate the maturation-dependent expression of the protein in leukocytes.
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