Adaptor and clathrin exchange at the plasma membrane and trans-Golgi network.
Adaptor and clathrin exchange at the plasma membrane and trans-Golgi network.
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DOI：
10.1091/mbc.e02-06-0353
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发表时间：
2003-02
影响因子：
3.3
通讯作者：
Xufeng Wu;Xiaohong Zhao;R. Puertollano;J. Bonifacino;E. Eisenberg;L. Greene
中科院分区：
文献类型：
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作者：
Xufeng Wu;Xiaohong Zhao;R. Puertollano;J. Bonifacino;E. Eisenberg;L. Greene
我们以前证明，使用荧光恢复后，光漂白，网格蛋白在网格蛋白包被的坑在质膜交换与游离网格蛋白在胞质溶胶中，这表明网格蛋白包被的坑是动态结构。我们现在调查是否网格蛋白在trans-Golgi网络以及网格蛋白适配器AP 2和AP 1网格蛋白包被的坑在质膜和trans-Golgi网络，分别也交换与游离蛋白质在胞质溶胶。我们发现，当网格蛋白包被的囊泡的出芽被阻止而不显着影响网格蛋白包被的凹坑的结构，网格蛋白和AP 2在质膜和网格蛋白和AP 1在trans-Golgi网络交换迅速与游离蛋白质在胞质溶胶中。相反，当网格蛋白包被的囊泡的出芽在质膜或trans-Golgi网络处被高渗蔗糖或K（+）耗尽阻断时，显著影响网格蛋白包被的凹坑的结构的条件，网格蛋白交换被阻断，但质膜处的AP 2以及trans-Golgi网络处的AP 1和GGA 1适配器继续快速交换。我们的结论是网格蛋白涂层坑是动态结构与快速交换网格蛋白和适配器，适配器能够交换独立的网格蛋白时，网格蛋白交换被阻止。
We previously demonstrated, using fluorescence recovery after photobleaching, that clathrin in clathrin-coated pits at the plasma membrane exchanges with free clathrin in the cytosol, suggesting that clathrin-coated pits are dynamic structures. We now investigated whether clathrin at the trans-Golgi network as well as the clathrin adaptors AP2 and AP1 in clathrin-coated pits at the plasma membrane and trans-Golgi network, respectively, also exchange with free proteins in the cytosol. We found that when the budding of clathrin-coated vesicle is blocked without significantly affecting the structure of clathrin-coated pits, both clathrin and AP2 at the plasma membrane and clathrin and AP1 at the trans-Golgi network exchange rapidly with free proteins in the cytosol. In contrast, when budding of clathrin-coated vesicles was blocked at the plasma membrane or trans-Golgi network by hypertonic sucrose or K(+) depletion, conditions that markedly affect the structure of clathrin-coated pits, clathrin exchange was blocked but AP2 at the plasma membrane and both AP1 and the GGA1 adaptor at the trans-Golgi network continue to rapidly exchange. We conclude that clathrin-coated pits are dynamic structures with rapid exchange of both clathrin and adaptors and that adaptors are able to exchange independently of clathrin when clathrin exchange is blocked.