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.

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+> - 和磷脂依赖性方式。其中，240K和80K蛋白分别确定为Calspectin和Synapsin I。在本地状态也观察到了膜联蛋白VI与这些蛋白质的结合。膜联蛋白VI在Ca^<2+>存在下与Calspectin的Calspectin结合Calspectin和Calspectin的Ps.An Annexin vi结合位点在Calspectin的存在下抑制了Calspectin和F-肌动蛋白之间的相互作用，位于肌动蛋白结合位点附近的B-Calspectin的N末端区域。另一方面，与cAMP依赖性蛋白激酶对突触I的磷酸化或CA^<2+>，钙调蛋白依赖性蛋白激酶的磷酸化抑制了与突触的N末端头部结合的膜联蛋白VI。 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.