| Project/Area Number |
03670142
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Kobe University |
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Principal Investigator |
KIKKAWA Ushio Kobe University Biosignal Research Center Professor, バイオシグナル研究センター, 教授 (40150354)
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| Co-Investigator(Kenkyū-buntansha) |
OGITA Kouji Kobe University School of Medicine Assistant, 医学部, 助手 (60204103)
NAKAMURA Shun-ichi Kobe University School of Medicine Associate Professor, 医学部, 助教授 (40155833)
浅岡 良則 神戸大学, バイオシグナル研究センター, 助手 (20222565)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1992: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Protein Kinase C / Signal Transduction / Protein Phosphorylation / Diacylglycerol / プロテインキナ-ゼC / ジアシルグリセロ-ル |
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| Research Abstract |
Protein kinase C plays crucial roles in signal transduction, and is a family of multiple subspecies. This study aimed to investigate the roles of the delta-, epsilon-, and zeta-subspecies, that are initially isolated from brain libraries by the analysis of cDNA clones. The delta-and epsilon-subspecies were purified to homogeneity from rat brain and shown to be dependent on diacylglycerol and phospholipid, but do not require calcium ion for their kinase activities. In addition, these two subspecies showed distinct enzymatic properties, the activity of the epsilon-subspecies is enhanced by fatty acids, whereas the delta-subspecies is inhibited. CHO cell lines overproducing each protein kinase C subspecies were established to study the effects of this enzyme family on cell phenotypes. The growth of the cells overproducing the delta-subspecies was markedly inhibited upon treatment with phorbol 12-myristate 13-acetate that is a potent activator of the protein kinase C family, whereas the cell lines expressing other subspecies were not significantly affected. These results suggest that each subspecies of protein kinase C family has distinct roles in growth regulation. As it was difficult to purify the zeta-subspecies form rat tissues, this subspecies was expressed with the baculovirus system and will be purified from the extract of the infected Sf9 cells. The delta- and epsilon-subspecies were also expressed in the insect cells. We are planning to prepare polyclonal antibodies against these subspecies to investigate the roles of the protein kinase C family by immunological procedures.
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