Studies on heterogeneity of protein kinase C, their structures, regulations and functions.

蛋白激酶C异质性及其结构、调控和功能的研究。

• 批准号: 63480126
• 负责人: KISHIMOTO Akira
• 金额: $ 5.76万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480126/
• 关键词: protein kinase C; cell proliferation; cell differentiation; diacylglycerol; phorbol ester; Ca^<2+>; inositol phospholipid; signal transduction; シアシルグリセロ-ル; 蛋白質リン酸化反応; プロテインキナーゼC; カルシウム; イノシトールリン脂質; ホルボールエステル; 蛋白質限定分解反応; カルパイン; ダウンレギュレーション

Protein kinase C (PKC) is a multifunctional protein Ser/Thr kinase and generally accepted to be involved in a wide variety of cellular signal transduction. In this study, we have found the existence of multiple subspecies of PKC through molecular cloning and enzymological analysis. Their relative activity and individual pattern of expression and intracellular locarization in several tissues and cell types are distinctly different one another. The PKC subspecies thus far obtained from various tissues exhibit subtle differences in their mode of activation, enzymological properties, and phorbol ester-induced down-regulation.In addition, the expressions of PKC subspecies are differently regulated during the differentiation of HL-60 cell toward granulocyte. Thus, it is strongly suggested that the members of PKC family each show distinctly different preference for substrate proteins that are located in specific intracellular comportments of the cell types in which they are expressed. Analysis of the activation of T-lymphocyte through PKC activation using phorbol esters or membrane-permeable diacylglycerols indicates a dual action of PKC, modulation of membrane receptor and regulation of gene expression. Short term PKC activation, which may be induced by the breakdown of inositol phospholipid, provides the down-regulation of TCR-CD3 complex on the surface of lymphocyte. On the other hand, sustained PKC activation is necessary to enhance the expression of IL-2 receptor eventually leading to cell proliferation.There appear to be several additional routes, other than the hydrolysis of inositol phospholipid, to provide the diacylglycerol that is needed for prolonged enzyme activation. This novel mechanisms leading to PKC activation as well as functional specificities of PKC subspecies remain to be explored to elucidate roles of PKC in the regulation of gene expression and growth control.

蛋白激酶C （PKC）是一种多功能蛋白丝氨酸/苏氨酸激酶，被广泛认为参与多种细胞信号转导。本研究通过分子克隆和酶学分析发现PKC存在多个亚种。它们在不同组织和细胞类型中的相对活性、个体表达模式和细胞内定位明显不同。迄今为止，从不同组织中获得的PKC亚种在其激活模式、酶学特性和磷酯诱导的下调方面表现出细微的差异。另外，在HL-60细胞向粒细胞分化的过程中，PKC亚种的表达受到不同程度的调控。因此，我们强烈建议PKC家族的每个成员对位于其表达的细胞类型的特定细胞内组分中的底物蛋白表现出明显不同的偏好。通过磷酸酯或膜透性二酰基甘油激活PKC对t淋巴细胞的激活分析表明PKC具有双重作用，即调节膜受体和调节基因表达。PKC的短期激活可能由肌醇磷脂的分解引起，从而下调淋巴细胞表面的TCR-CD3复合物。另一方面，PKC持续激活是增强IL-2受体表达最终导致细胞增殖的必要条件。除了肌醇磷脂的水解外，似乎还有几种其他途径可以提供延长酶激活所需的二酰基甘油。这种导致PKC激活的新机制以及PKC亚种的功能特异性仍有待探索，以阐明PKC在基因表达调控和生长控制中的作用。

项目成果

Kishimoto,A.: "The protein kinase C family in the brain:heterogeneity and its implications." Ann.N.Y.Acad.Sci.(in press). 568. (1990)

Kishimoto,A.：“大脑中的蛋白激酶 C 家族：异质性及其影响。”

Ogita,K.: "The heterogeneity of protein kinase C in signal transduction cascade." Adv.Second Messenger Phosphoprotein Res.(in press). (1990)

Ogita,K.：“信号转导级联中蛋白激酶 C 的异质性。”

Ogita, K., Kikkawa, U., Shearman, M.S., Ase, K., Berry, N., Kishimoto, A., and Nishizuka, Y.: "The structure and function of multispecies of protein kinase C." Adv. Prostaglandin Thromboxane Leukotriene Res. 19, 49-56 (1989).

Ogita, K.、Kikkawa, U.、Shearman, M.S.、Ase, K.、Berry, N.、Kishimoto, A. 和 Nishizuka, Y.：“多种蛋白激酶 C 的结构和功能。”

Kikkawa, U., Kishimoto, A., and Nishizuka, Y.: "The protein kinase C family: heterogeneity and its implications." Annu. Rev. Biochem. 58, 31-44 (1989).

Kikkawa, U.、Kishimoto, A. 和 Nishizuka, Y.：“蛋白激酶 C 家族：异质性及其影响。”

Shearman, M.S., Naor, Z., Sekiguchi, K., Kishimoto, A., and Nishizuka, Y.: "Selective activation of the gamma-subspecies of protein kinase C from bovine cerebellum by arachidonic acid and its lipoxygenase metabolites." FEBS Lett. 243, 177-182 (1989).

Shearman, M.S.、Naor, Z.、Sekiguchi, K.、Kishimoto, A. 和 Nishizuka, Y.：“花生四烯酸及其脂氧合酶代谢物选择性激活牛小脑蛋白激酶 C 的 γ 亚种。”