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Dynamic physiological roles of calcium binding proteins in the brain.

钙结合蛋白在大脑中的动态生理作用。

基本信息

批准号：
01044105
负责人：
HATASE Osamu
金额：
$ 4.74万
依托单位：
The Kagawa Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01044105/
关键词：
Calcium Binding Protein Calcineurin Calmodulin Lipocortin カルシニュウリン

项目摘要

Three Ca^<2+>-binding proteins in brain, calcineurin, phosphodiesterase and lipocortins, were studied and following results were obtained.l. calcineurin (CaN)・Monoclonal antibodies against alpha and beta were produced.・Immunohistochemical study elucidated the distinct distribution of both subunits in brain.・A novel beta subunit (beta_1) from rat testis was cloned and amino acid sequence was determined. Six additive hydrophilic amino acids were located in the carboxyl terminal of the molecule as compared to the bovine type beta subunit.・Two isoforms of alpha subunit of bovine brain CaN were demonstrated by Western blot analysis using the monoclonal antibody.・A part of CaN could translocate from cytosoi to membrane in a Ca^<2+> dependent manner.2. phosphodiesterase (PDE)・A monclonal antibody was raised.・Two isozymes, PDE_1, (Mr=63, 000) and PDE_2 (Mr=60, 000), were identified.・PDE_1 and PDE_2 were phosphorylated by protein kinase C and cyclic-AMP dependent protein kinase, respectively.・PDE_1 and PDE_2 showed a distinct distribution in rat brain.3. lipocortins (LP)・Monoclonal antibodies against LP-I (Mr=35, 000) and LP-II (Mr=36, 000) were produced.・Both LP-I and LP-II inhibited phospholipase A_2 activity, and phosphorylated forms of both proteins by protein kinase C showed a reduced inhibitory activity on phospholipase A_2.・LP-I and LP-11 showed a developmental change in amount ; higher quantity in newborn rat brain and lower in adult brain.・Specific distribution in astrocytes (not in neurons) was confirmed.

研究了脑中的三种Ca 2+ 结合蛋白：钙调神经磷酸酶、磷酸二酯酶和脂皮质素，并获得了以下结果。钙调神经磷酸酶 (CaN) ・产生了针对 α 和 β 的单克隆抗体。・免疫组织化学研究阐明了这两个亚基在脑中的独特分布。・从大鼠睾丸中克隆了一种新的 β 亚基 (beta_1)，并确定了氨基酸序列。与牛型β亚基相比，分子的羧基末端有6个附加的亲水性氨基酸。・使用单克隆抗体进行蛋白质印迹分析，证实了牛脑CaN的α亚基的两种亚型。・CaN的一部分可以以Ca^2+依赖性方式从胞浆易位到膜。 2．磷酸二酯酶 (PDE) ・产生单克隆抗体。・鉴定出 PDE_1, (Mr=63, 000) 和 PDE_2 (Mr=60, 000) 两种同工酶。・PDE_1 和 PDE_2 分别被蛋白激酶 C 和环 AMP 依赖性蛋白激酶磷酸化。・PDE_1 和 PDE_2 PDE_2在大鼠脑内有明显的分布。 3．脂皮质素 (LP) ・生产针对 LP-I (Mr=35, 000) 和 LP-II (Mr=36, 000) 的单克隆抗体。・LP-I 和 LP-II 均抑制磷脂酶 A_2 活性，并且蛋白激酶 C 将两种蛋白磷酸化后，磷脂酶抑制活性降低 A_2.·LP-I和LP-11在数量上表现出发育变化；新生大鼠脑中的含量较高，成年大鼠脑中的含量较低。・已确认星形胶质细胞（而非神经元）中的特定分布。