| Project/Area Number |
01440027
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
HIDAKA Hiroyoshi Nagoya Univ., School of Med., professor, 医学部, 教授 (80100171)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Ryoji Nagoya Univ., School of Med., associate professor, 医学部, 助教授 (00020917)
HAGIWARA Masatoshi Nagoya Univ., School of Med., assistant professor, 医学部, 助手 (10208423)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥19,300,000 (Direct Cost: ¥19,300,000)
Fiscal Year 1991: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1990: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1989: ¥12,200,000 (Direct Cost: ¥12,200,000)
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| Keywords | Protein phosphorylation / Ca^<2+>-dependent protein kinase / CaM-kinase II / Protein kinase C / EIA / KN-62 / H-89 / Protein kinase inhibitor / Aーキナ-ゼ / δ型Cキナ-ゼ / ζ型Cキナ-ゼ / 長期増強現象 / GABA放出 / ACAMPー81 / MARCKS / Cキナ-ゼアイソザイム / エンザイムイムノアッセイ / Cキナ-ゼアイソザイムの細胞特異的発現 / 特異的阻害剤 |
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| Research Abstract |
To elucidate the functions of Ca^<2+>-dependent protein kinases in central nervous system in health and diseases, biochemical and pharmacological approaches were performed in this study. First, we have developed the subtype specific enzyme immunoassay (EIA) using three anti-protein kinase C (PKC) isozymes monoclonal antibodies. The EIA showed the quantitative analysis and subtype-specific expression of PKC in CNS tumor cells including glioblastoma and neuroblastoma cells. The results suggested that PKC isozymes have a cell-type specificity in CNS tumors, and that detection of PKC isozymes will aid in understanding the diverse effects of PKC and also to diagnose tumors derived. from glia or neuronal
cells. In addition, we produced antibodies against the novel PKCS, and examined tissue and cell-type specific expression of the novel PKCS.
Second, we have newly synthesized specific inhibitors of various protein ldnases including Ca^<2+>/calmodulin protein kinase II (CaM Idnase 11), cAMp-dependent protein kinase (A-kinase) and casein kidase I. KN-62 is specific CaM kinase 11 inhibitor and dose not inhibit other kinase activities. Furtennore, KN-62 inhibit the releasing of GABA and long term potentiation in central nervous system suggesting that the compound is a useful tool for elucidation of the functions of CaM kinase H in many tissues. H-89 (A-kinase inhibitor) also inhibited neurite growth n PC- 12 cells induced by folskolin, and induction of immediately early gene.
We could determined the cell-type specific expression of PKC isozymes and developed specific inhibitors of various protein kinases. We should try on next steps to elucidate the functions of these protein Idnases using specific inhibitors.
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