Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network
Protein kinase A activity is required for the budding of constitutive transport vesicles from the trans-Golgi network
复制标题
DOI：
10.1073/pnas.94.26.14461
复制
发表时间：
1997-12-23
影响因子：
11.1
通讯作者：
Velasco, A
中科院分区：
文献类型：
--
作者：
Muñiz, M;Martín, ME;Velasco, A
我们研究了蛋白激酶A（PKA）在囊泡介导的蛋白质从高尔基体网络（TGN）转运到细胞表面的作用。在体内，该转运步骤被PKA催化亚基（C-PKA）的抑制剂抑制，例如被称为H89的化合物和热稳定PKA抑制剂中所含的抑制肽序列的肉豆蔻酰化形式。H89的抑制作用发生在水泡性口炎病毒G糖蛋白从TGN转移到细胞表面的早期阶段。从这种抑制相关的一个短暂的增加，在高尔基体区的自由包被囊泡的数量referenced。使用水泡性口炎病毒感染的透化细胞在体外研究了TGN的水泡出芽。加入C-PKA刺激囊泡释放，而PKA抑制肽H89和抗C-PKA抗体抑制囊泡释放。此外，囊泡释放减少时，PKA耗尽的胞质溶胶中使用和恢复添加C-PKA。这些结果表明，PKA活性在从TGN的组成性运输囊泡的生产中的调节作用。
We have examined the role played by protein kinase A (PKA) in vesicle-mediated protein transport from the trans-Golgi network (TGN) to the cell surface. In vivo this transport step,vas inhibited by inhibitors of PKA catalytic subunits (C-PKA) such as the compound known as H89 and a myristoylated form of the inhibitory peptide sequence contained in the thermostable PKA inhibitor. Inhibition by H89 occurred at an early stage during the transfer of vesicular stomatitis virus G glycoprotein from the TGN to the cell surface. Reversal from this inhibition correlated with a transient increase in the number of free coated vesicles in the Golgi area. Vesicle budding from the TGN was studied in vitro using vesicular stomatitis virus-infected, permeabilized cells. Addition to this assay of C-PKA stimulated vesicle release while it was suppressed by PKA inhibitory peptide, H89, and antibody against C-PKA. Furthermore, vesicle release was decreased when PKA-depleted cytosol was used and restored by addition of C-PKA. These results indicate a regulatory role for PKA activity in the production of constitutive transport vesicles from the TGN.