权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Regulation of cytoskeleton by annexin VI

膜联蛋白 VI 对细胞骨架的调节

基本信息

批准号：
07680842
负责人：
INUI Makoto
金额：
$ 1.41万
依托单位：
Yamaguchi University (1996)Osaka University (1995)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680842/
关键词：
Annexin Calspectin Synapsin I Calcium ion Phospholipid Phosphorylation

项目摘要

To elucidate the physiological significance of annexin VI in brain, we developed a method to detect annexin VI-binding proteins using ^<125> I-annexin VI on a nitrocellulose sheet to which rat brain proteins were electrophoretically transferred after SDS polyacrylamide gel electrophoresis.We found several annexin VI-binding proteins, which interact with annexin VI in a Ca^<2+>- and phospholipid-dependent manner. Of these, the 240K and the 80K proteins were identified to be calspectin and synapsin I,respectively. The bindings of annexin VI to these proteins were also observed in the native state. Annexin VI inhibited the interaction between calspectin and F-actin by binding to calspectin in the presence of Ca^<2+> and PS.An annexin VI-binding site in calspectin was localized to the N-terminal region of b-calspectin near the actin-binding site. On the other hand, annexin VI bound to the N-terminal head region of synapsin I.The binding was inhibited by phosphorylation of synapsin I by cAMP-dependent protein kinase or by Ca^<2+>, calmodulin-dependent protein kinase II.We further examined the direct interaction between these two annexin VI-binding proteins. The interaction of these proteins was high affinity one with Kd of about 10 mM.The binding was inhibited by phosphorylation of synapsin I by cAMP-dependent protein kinase or by Ca^<2+>, calmodulin-dependent protein kinase II.These results indicate that annexin VI-binding proteins play an important role in the regulation of neurotransmitter release in the presynaptic region.

为了阐明膜联蛋白 VI 在大脑中的生理意义，我们开发了一种使用 ^<125> I-膜联蛋白 VI 检测膜联蛋白 VI 结合蛋白的方法，该方法使用硝酸纤维素片上的 ^<125> I-膜联蛋白 VI，在 SDS 聚丙烯酰胺凝胶电泳后将大鼠脑蛋白电泳转移到硝化纤维素片上。我们发现了几种膜联蛋白 VI 结合蛋白，它们与膜联蛋白 VI 相互作用。 Ca 2+ -和磷脂依赖性方式。其中，240K 和 80K 蛋白分别被鉴定为 calspectin 和突触蛋白 I。在天然状态下也观察到膜联蛋白 VI 与这些蛋白质的结合。膜联蛋白VI通过在Ca 2+ 和PS存在下与钙观蛋白结合来抑制钙观蛋白和F-肌动蛋白之间的相互作用。钙观蛋白中的膜联蛋白VI结合位点位于肌动蛋白结合位点附近的b-钙观蛋白的N端区域。另一方面，膜联蛋白VI与突触蛋白I的N末端头部区域结合。突触蛋白I的磷酸化被cAMP依赖性蛋白激酶或钙调蛋白依赖性蛋白激酶II的Ca^2+抑制。我们进一步检查了这两种膜联蛋白VI结合蛋白之间的直接相互作用。这些蛋白质的相互作用是高亲和力的，Kd约为10 mM。这种结合被cAMP依赖性蛋白激酶或钙调蛋白依赖性蛋白激酶II的Ca^2+磷酸化突触蛋白I所抑制。这些结果表明膜联蛋白VI结合蛋白在调节突触前区神经递质释放中发挥重要作用。