1990 Fiscal Year Annual Research Report

Protein Kinase Cによる情報伝達の機構

Research Project

Project/Area Number	01065006
Research Institution	Kobe University
Principal Investigator	西塚泰美神戸大学, 医学部, 教授 (10025546)
Co-Investigator(Kenkyū-buntansha)	岸本明神戸大学, 医学部, 助教授 (60127363)
Keywords	プロテインキナ-ゼC / 細胞内情報伝達 / カルシウムイオン / イノシト-ルリン脂質 / ジアシルグリセロ-ル / ホルボ-ルエステル / 細胞増殖 / 細胞分化
Research Abstract	Protein Kinase C(PKC)は多彩な分子種からなる酵素群であり、外界情報の受容伝達の主軸として、多岐の生理活性物質による細胞機能の調節に深く関与していることが確実視されている。本研究ではPKCを介する情報伝達機構の化学的基盤と生理を解析し、病態分析等に応用することを目的としている。当初2カ年の研究計画は順調に進行しており、PKC分子種の多様性の酵素化学等の基礎的研究はほぼ予定どおり達成されている。本年度では基礎的成果に立脚してPKCの機能の解析へと重点を移し、ことに神経機能解明の緒を開くことができた。昨年度の解析から、記憶中枢である海馬領域のプレシナプスではα、β種PKCが存在し、γ種が存在しない。実際に種々のPKCを有するシナプトソ-ムを調整し、高K^+等の刺激を行った所、ノルエピネフリンの放出反応にはα種とβ種のPKCが関与しており、γ種は無関係であることが示された。このα、β種PKC活性化においてアラキドン酸やドコサヘキサエノイン酸等は、ジアシルグリセロ-ルによるPKCの活性化を相乗的に高めることから、PKCがそれらの不飽和脂肪酸の作用点の少なくとも一つであることが判明した。これら神経系において生物活性を持つ不飽和脂肪酸のプレシナプスでの作用機作、特にLTP(Long Term Potentiation)における具体的役割を示唆する最初の知見となっている。一方、Tリンパ球の活性化はCa^<2+>のシグナルとPKCのシグナルの協同作用で行われると考えられる。この時、持続的なジアシルグリセロ-ルの上昇が必要であり、PKCの恒常的な活性化が細胞の増殖に関与することが判明した。上述の様にLTP等の神経機能においてもPKCの持続的な活性化が問題となっており、PKCのシグナルの解明にはジアルグリセロ-ル、不飽和脂肪酸の動向の分析が必須なものとなっている。特異的基質を介した他のシグナルとのcrossーtalk、ことにcyclic AMPを介するシグナル、受容体を介入したチロシンキナ-ゼがジアシルグリセロ-ル、不飽和脂肪酸産生とどのように関係しているかが次の大きな課題となっている。

Research Products

(21 results)

All Other

All Publications (21 results)

[Publications] Nishizuka,Y.: "Protein Kinase C family and nervous function." Progress Brain Res.(1991)
[Publications] Berry,N.: "Protein Kinase C and T cell activation." Eur.J.Biochem.189. 205-214 (1990)
[Publications] Farago,A.: "Protein Kinase C in transmembrane signalling." FEBS Lett.268. 350-354 (1990)
[Publications] Ogita,K.: "The heterogeneity of protein kinase C in signal transduction cascade." Adv.Second Messenger and Phosphoprotein Res.24. 218-223 (1990)
[Publications] Ishii,S.: "Structure and function of the protein CREーBP1:heterogeneity of the protein binds to the cAMPーresponsive element." Adv.Second Messenger and Phosphoprotein Res.24. 335-339 (1990)
[Publications] Kishimoto,A.: "Limited proteolysis of protein kinase C by calpain,its possible implication." Adv.Second Messenger and Phosphoprotein Res.24. 472-477 (1990)
[Publications] Yasuda,I.: "A synthetic peptide substrate for selective assay of protein kinase C." Biochem.Biophys.Res.Commun.166. 1220-1227 (1990)
[Publications] Bignon,E.: "Dual action of hydroxylated diphenylethylenes on protein kinase C." Biochem.Biophys.Res.Commun.166. 1471-1478 (1990)
[Publications] Bignon,E.: "Protein Kinase C subspecies in estrogen receptorーpositive and ーnegative human breast cancer cell lines." Biochem.Biophys.Res.Commun.171. 1071-1078 (1990)
[Publications] Hashimoto,K.: "Protein Kinase C during differentiation of human promyelocytic leukemia cell line HLー60." FEBS Lett.263. 31-34 (1990)
[Publications] Bignon,E.: "Multiple mechanisms of protein kinase C inhibition by triphenylacrylonitrile antiestrogens." FEBS Lett.271. 54-58 (1990)
[Publications] Shearman,M.S.: "Protein Kinase C Subspecies in adult rat hippocampal symaptosomes;activation by diacylglycerol and arachidonic acid." FEBS Lett.(1991)
[Publications] Berry,N.: "Activation of resting human T cells requires prolonged stimulation of protein kinase C." Proc.Natl.Acad.Sci.87. 2294-2298 (1990)
[Publications] Ito,A.: "Immunocytochemical localization of α subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.87. 3195-3199 (1990)
[Publications] Ogita,K.: "Protein Kinase C in Saccharomyces cerevisiae:comparison with the mammalian enzyme." Proc.Natl.Acad.Sci.87. 5011-5015 (1990)
[Publications] Shinomura,T.: "Synergistic action of diacylglycerol and unsaturated fatty acid for protein kinase C activation,its possible implications." Proc.Natl.Acad.Sci.(1991)
[Publications] Shearman,M.S.: "Synaptosomal protein kinase C subspecies;dynamic change in the hippocampus and cerebellar cortex concomitant with synaptogenesis." J.Neurochem.(1991)
[Publications] Oda,T.: "Synaptosomal protein kinase C subspecies;downーregulation promoted by phorbol ester and its effect on evoked norepinephrine release." J.Neurochem.(1991)
[Publications] Ogita,K.: "Expression,separation,and assay of protein kinase C subspecies." Methods enzymol.(1991)
[Publications] Kishimoto,A.: "Preparation of protein Kinase C subspeciesーspecific antiーpeptid antibodies for immunostaining." Methods enzymol.(1991)
[Publications] Kishimoto,A.: "Synthesis of peptide substrates for protein kinase C." Methods Neurosci.(1991)

1990 Fiscal Year Annual Research Report

Protein Kinase Cによる情報伝達の機構

Principal Investigator

西塚 泰美 神戸大学, 医学部, 教授 (10025546)

Research Products

[Publications] Nishizuka,Y.: "Protein Kinase C family and nervous function." Progress Brain Res.(1991)

[Publications] Berry,N.: "Protein Kinase C and T cell activation." Eur.J.Biochem.189. 205-214 (1990)

[Publications] Farago,A.: "Protein Kinase C in transmembrane signalling." FEBS Lett.268. 350-354 (1990)

[Publications] Ogita,K.: "The heterogeneity of protein kinase C in signal transduction cascade." Adv.Second Messenger and Phosphoprotein Res.24. 218-223 (1990)

[Publications] Ishii,S.: "Structure and function of the protein CREーBP1:heterogeneity of the protein binds to the cAMPーresponsive element." Adv.Second Messenger and Phosphoprotein Res.24. 335-339 (1990)

[Publications] Kishimoto,A.: "Limited proteolysis of protein kinase C by calpain,its possible implication." Adv.Second Messenger and Phosphoprotein Res.24. 472-477 (1990)

[Publications] Yasuda,I.: "A synthetic peptide substrate for selective assay of protein kinase C." Biochem.Biophys.Res.Commun.166. 1220-1227 (1990)

[Publications] Bignon,E.: "Dual action of hydroxylated diphenylethylenes on protein kinase C." Biochem.Biophys.Res.Commun.166. 1471-1478 (1990)

[Publications] Bignon,E.: "Protein Kinase C subspecies in estrogen receptorーpositive and ーnegative human breast cancer cell lines." Biochem.Biophys.Res.Commun.171. 1071-1078 (1990)

[Publications] Hashimoto,K.: "Protein Kinase C during differentiation of human promyelocytic leukemia cell line HLー60." FEBS Lett.263. 31-34 (1990)

[Publications] Bignon,E.: "Multiple mechanisms of protein kinase C inhibition by triphenylacrylonitrile antiestrogens." FEBS Lett.271. 54-58 (1990)

[Publications] Shearman,M.S.: "Protein Kinase C Subspecies in adult rat hippocampal symaptosomes;activation by diacylglycerol and arachidonic acid." FEBS Lett.(1991)

[Publications] Berry,N.: "Activation of resting human T cells requires prolonged stimulation of protein kinase C." Proc.Natl.Acad.Sci.87. 2294-2298 (1990)

[Publications] Ito,A.: "Immunocytochemical localization of α subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.87. 3195-3199 (1990)

[Publications] Ogita,K.: "Protein Kinase C in Saccharomyces cerevisiae:comparison with the mammalian enzyme." Proc.Natl.Acad.Sci.87. 5011-5015 (1990)

[Publications] Shinomura,T.: "Synergistic action of diacylglycerol and unsaturated fatty acid for protein kinase C activation,its possible implications." Proc.Natl.Acad.Sci.(1991)

[Publications] Shearman,M.S.: "Synaptosomal protein kinase C subspecies;dynamic change in the hippocampus and cerebellar cortex concomitant with synaptogenesis." J.Neurochem.(1991)

[Publications] Oda,T.: "Synaptosomal protein kinase C subspecies;downーregulation promoted by phorbol ester and its effect on evoked norepinephrine release." J.Neurochem.(1991)

[Publications] Ogita,K.: "Expression,separation,and assay of protein kinase C subspecies." Methods enzymol.(1991)

[Publications] Kishimoto,A.: "Preparation of protein Kinase C subspeciesーspecific antiーpeptid antibodies for immunostaining." Methods enzymol.(1991)

[Publications] Kishimoto,A.: "Synthesis of peptide substrates for protein kinase C." Methods Neurosci.(1991)

西塚泰美神戸大学, 医学部, 教授 (10025546)