1989 年度実績報告書

Protein Kinase Cによる情報伝達の機構

研究課題

研究課題/領域番号	01065006
研究機関	神戸大学
研究代表者	西塚泰美神戸大学, 医学部, 教授 (10025546)
研究分担者	齋藤尚亮神戸大学, 医学部, 助手 (60178499) 岸本明神戸大学, 医学部, 助教授 (60127363)
キーワード	プロテインキナーゼC / 細胞内情報伝達 / カルシウムイオン / イノシトールリン脂質 / ジアシルグリセロール / ホルボールエステル / 細胞増殖 / 細胞分化
研究概要	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。初年度の基礎的研究は順調に進行し、初期の成果を得た。特に、遺伝子工学的手法によって特異的分子種を安定して発現する細胞系を確立し、生理機能の解明へとつなげている。また、PKCのホルボールエステル結合領域を決定した。これはPKCの活性化機作の解明、ことに脂質と蛋白質の相互識別の課題に新しい視点を開くものである。一方、電顕レベルでの細胞内局在の詳細な解析によっても細胞膜やゴルジ小体にのみ局在する分子種を明らかにし、それぞれの特異的機能の解明に重要な示唆を与えている。これらの基礎的成果に加えて、次年度に向けてPKCの具体的機能の解明にも着手した。すなわち、Tーリンパ球、HLー60細胞や大脳シナプトソームを用いて外界シグナルによる活性化、分化、増殖における分子種の特異的動態やダウンレギュレーションの機構を明らかにしつつある。ことに、Tリンパ球の増殖、活性化には短期間のPKCの活性化による細胞膜受容体の調節と長期間のPKCの活性化による遺伝子発現の調節の二種類の作用メカニズムが働いている可能性が高い。この結果は、従来のイノシトールリン脂質の代謝回転による短期間のジアシルグリセロールの産生に対する応答のみでなく、長期間のジアシルグリセロール産生機序の解明とその間のPKC分子種に特異的な活性化や動態の把握という新しい課題を与えるものである。また、CRE結合蛋白質がPKCの標的蛋白質であることが決定的となっており、遺伝子発現の調節におけるcyclic AMP系やtyrosin kinase系との共同作用の解明へとつなげることができた。

研究成果
(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] Kishimoto,A.: "Limited proteolysis of protein kinase C subspecies by calcium-dependent neutral protease(calpain)." J.Biol.Chem.264. 4088-4092 (1989)
[文献書誌] Shearman,M.S.: "Selective activation of the γ-subspecies of protein kinase C from bovine cerebellum by arachidonic acid and its lipoxygenase metabolites." FEBS Lett.243. 177-182 (1989)
[文献書誌] Sawamura,S.: "Expression of protein kinase C subspecies in human leukemia-lymphoma cell lines." FEBS Lett.247. 353-357 (1989)
[文献書誌] Hosoda,K.: "Immunocytochemical localization of the β1 subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. 86. 1393-1397 (1989)
[文献書誌] Saito,N.: "Immunocytochemical localization of the β2 subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. 86. 3409-3413 (1989)
[文献書誌] Ono,Y.: "Protein kinase C ζ subspecies from rat brain:Its structure,expression and properties." Proc.Natl.Acad.Sci.USA. 86. 3099-3103 (1989)
[文献書誌] Nishizuka,Y.: "Studies and prospective of the protein kinase C family for cellular regulation." Cancer. 63. 1892-1903 (1989)
[文献書誌] Nishizuka,Y.: "The family of protein kinase C for signal transduction." J.Am.Med.Assoc.262. 1826-1833 (1989)
[文献書誌] Kajikawa,N.: "Membrane-permeable diacylglycerol,its application to platelet secretion,and regulation of platelet protein kinase C." Methods Enzymol.169. 430-442 (1989)
[文献書誌] Kikkawa,U.: "The heterogeneity of protein kinase C and its implications for neuronal cell regulation." Brain Signal Transduction and Memory. 21-32 (1989)
[文献書誌] Yasuda,I.: "A synthetic peptide substrate for selective assay of protein kinase C." Biochem.Biophys.Res.Commun.166. 1220-1227 (1990)
[文献書誌] Bignon,E.: "Dual action of hydroxylated diphenylethylenes on protein kinase C." Biochem.Biophys.Res.Commun.(1990)
[文献書誌] Ono,Y.: "Phorbol ester binding to protein kinase C requires cysteine-rich zinc-finger-like sequence." Proc.Natl.Acad.Sci.USA. 86. 4868-4871 (1989)
[文献書誌] Berry,N.: "Human T cell activation by phorbol esters and diacylglycerol analogues." J.Immunol.143. 1407-1413 (1989)
[文献書誌] Bignon,E.: "Modes of inhibition of protein kinase C by triphenylacrylonitrile antiestrogens." Biochem.Biophys.Res.Commun.163. 1377-1383 (1989)
[文献書誌] Shearman,M.S.: "Modulation of ion channels;a key function of the protein kinase C family." Pharmacol.Reviews. 41. 211-237 (1989)
[文献書誌] Kikkawa,U.: "The protein kinase C family:heterogeneity and its implications." Ann.Rev.Biochem.58. 31-44 (1989)
[文献書誌] Ogita,K.: "The structure and function of multispecies of protein kinase C." Adv.in Pros.,Thrombozane and Leukotriene Res.19. 49-56 (1989)
[文献書誌] Ito,A.: "Immunocytochemical localization of α subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. (1990)
[文献書誌] Berry,N.: "Activation of resting human T cells requires prolonged stimulation of protein kinase C." Proc.Natl.Acad.Sci.USA. (1990)
[文献書誌] Hashimoto,K.: "Protein kinase C during differentiation of human promyelocytic leukemia cell line HL-60." FEBS Lett.(1990)
[文献書誌] Berry,N.: "Protein kinase C and T cell activation." Eur.J.Biochem.(1990)
[文献書誌] Kishimoto,A.: "The protein kinase C family in the brain:heterogeneity and its implications." Ann.N.Y.Acad.Sci.(1990)
[文献書誌] Ogita,K.: "The heterogeneity of protein kinase C in siganl transduction cascade." Adv.Second Messenger Phosphoprotein Res.(1990)
[文献書誌] Ishii,S.: "Structure and function of the protein CRE-BP1:heterogeneity of the protein that binds to the cAMP-responsive element." Adv.Second Messenger Phosphoprotein Res.(1990)
[文献書誌] Kishimoto,A.: "Limited proteolysis of protein kinase C by calpain,its possible implication." Adv.Second Messenger Phosphoprotein Res.(1990)
[文献書誌] Hashimoto,K.: "Calpains and regulation of protein kinase C." The Calcium-dependent Proteolytic System of Mammalian Cells.(1990)

1989 年度 実績報告書

Protein Kinase Cによる情報伝達の機構

研究代表者

西塚 泰美 神戸大学, 医学部, 教授 (10025546)

研究成果

[文献書誌] Kishimoto,A.: "Limited proteolysis of protein kinase C subspecies by calcium-dependent neutral protease(calpain)." J.Biol.Chem.264. 4088-4092 (1989)

[文献書誌] Shearman,M.S.: "Selective activation of the γ-subspecies of protein kinase C from bovine cerebellum by arachidonic acid and its lipoxygenase metabolites." FEBS Lett.243. 177-182 (1989)

[文献書誌] Sawamura,S.: "Expression of protein kinase C subspecies in human leukemia-lymphoma cell lines." FEBS Lett.247. 353-357 (1989)

[文献書誌] Hosoda,K.: "Immunocytochemical localization of the β1 subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. 86. 1393-1397 (1989)

[文献書誌] Saito,N.: "Immunocytochemical localization of the β2 subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. 86. 3409-3413 (1989)

[文献書誌] Ono,Y.: "Protein kinase C ζ subspecies from rat brain:Its structure,expression and properties." Proc.Natl.Acad.Sci.USA. 86. 3099-3103 (1989)

[文献書誌] Nishizuka,Y.: "Studies and prospective of the protein kinase C family for cellular regulation." Cancer. 63. 1892-1903 (1989)

[文献書誌] Nishizuka,Y.: "The family of protein kinase C for signal transduction." J.Am.Med.Assoc.262. 1826-1833 (1989)

[文献書誌] Kajikawa,N.: "Membrane-permeable diacylglycerol,its application to platelet secretion,and regulation of platelet protein kinase C." Methods Enzymol.169. 430-442 (1989)

[文献書誌] Kikkawa,U.: "The heterogeneity of protein kinase C and its implications for neuronal cell regulation." Brain Signal Transduction and Memory. 21-32 (1989)

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

[文献書誌] Bignon,E.: "Dual action of hydroxylated diphenylethylenes on protein kinase C." Biochem.Biophys.Res.Commun.(1990)

[文献書誌] Ono,Y.: "Phorbol ester binding to protein kinase C requires cysteine-rich zinc-finger-like sequence." Proc.Natl.Acad.Sci.USA. 86. 4868-4871 (1989)

[文献書誌] Berry,N.: "Human T cell activation by phorbol esters and diacylglycerol analogues." J.Immunol.143. 1407-1413 (1989)

[文献書誌] Bignon,E.: "Modes of inhibition of protein kinase C by triphenylacrylonitrile antiestrogens." Biochem.Biophys.Res.Commun.163. 1377-1383 (1989)

[文献書誌] Shearman,M.S.: "Modulation of ion channels;a key function of the protein kinase C family." Pharmacol.Reviews. 41. 211-237 (1989)

[文献書誌] Kikkawa,U.: "The protein kinase C family:heterogeneity and its implications." Ann.Rev.Biochem.58. 31-44 (1989)

[文献書誌] Ogita,K.: "The structure and function of multispecies of protein kinase C." Adv.in Pros.,Thrombozane and Leukotriene Res.19. 49-56 (1989)

[文献書誌] Ito,A.: "Immunocytochemical localization of α subspecies of protein kinase C in rat brain." Proc.Natl.Acad.Sci.USA. (1990)

[文献書誌] Berry,N.: "Activation of resting human T cells requires prolonged stimulation of protein kinase C." Proc.Natl.Acad.Sci.USA. (1990)

[文献書誌] Hashimoto,K.: "Protein kinase C during differentiation of human promyelocytic leukemia cell line HL-60." FEBS Lett.(1990)

[文献書誌] Berry,N.: "Protein kinase C and T cell activation." Eur.J.Biochem.(1990)

[文献書誌] Kishimoto,A.: "The protein kinase C family in the brain:heterogeneity and its implications." Ann.N.Y.Acad.Sci.(1990)

[文献書誌] Ogita,K.: "The heterogeneity of protein kinase C in siganl transduction cascade." Adv.Second Messenger Phosphoprotein Res.(1990)

[文献書誌] Ishii,S.: "Structure and function of the protein CRE-BP1:heterogeneity of the protein that binds to the cAMP-responsive element." Adv.Second Messenger Phosphoprotein Res.(1990)

[文献書誌] Kishimoto,A.: "Limited proteolysis of protein kinase C by calpain,its possible implication." Adv.Second Messenger Phosphoprotein Res.(1990)

[文献書誌] Hashimoto,K.: "Calpains and regulation of protein kinase C." The Calcium-dependent Proteolytic System of Mammalian Cells.(1990)

1989 年度実績報告書

西塚泰美神戸大学, 医学部, 教授 (10025546)