Regulation of CB1 cannabinoid receptor internalization by a promiscuous phosphorylation-dependent mechanism.
Regulation of CB1 cannabinoid receptor internalization by a promiscuous phosphorylation-dependent mechanism.
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通过滥交磷酸化依赖性机制调节CB1大麻素受体内在化。
DOI：
10.1111/j.1471-4159.2008.05336.x
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发表时间：
2008-07
影响因子：
4.7
通讯作者：
Mackie K
中科院分区：
文献类型：
--
作者：
Daigle TL;Kwok ML;Mackie K
激动剂刺激CB1受体内化。以前的工作表明，受体的极端羧基末端调节这种内化-可能是通过聚集在该区域内的丝氨酸和苏氨酸的磷酸化来实现的。虽然羧基末端(V460Z CB1)的截断和随后这些假定的磷酸化位点的移除阻止了AtT20细胞的内吞作用，但CB1受体内化所需的残基仍然难以捉摸。为了确定内化的结构要求，我们评估了在HEK293细胞中稳定表达的羧基末端突变CB1受体的内吞作用。与AtT20细胞相比，在HEK293细胞中表达的V460Z CB1受体内化的程度和动力学与野生型受体相似。然而，当这些受体在HEK293细胞中表达时，极端羧基末端的丝氨酸和/或苏氨酸残基的突变会减弱内化。这些结果表明，在HEK293细胞中，极端的羧基末端磷酸化位点不是截断受体内化所必需的，而是全长受体内化所必需的。βarrestin-2与突变的cb1受体结合的分析表明，推测的羧基末端磷酸化位点介导了βarrestin-2的易位。这项研究表明，局部细胞环境影响CB1受体内化的结构决定因素。此外，磷酸化可能调节(全长)CB1受体的内化。
Agonists stimulate CB1 receptor internalization. Previous work suggests that the extreme carboxy-terminus of the receptor regulates this internalization – likely through the phosphorylation of serines and threonines clustered within this region. While truncation of the carboxy-terminus (V460Z CB1) and consequent removal of these putative phosphorylation sites prevents endocytosis in AtT20 cells, the residues necessary for CB1 receptor internalization remain elusive. To determine the structural requirements for internalization, we evaluated endocytosis of carboxy-terminal mutant CB1 receptors stably expressed in HEK293 cells. In contrast to AtT20 cells, V460Z CB1 receptors expressed in HEK293 cells internalized to the same extent and with similar kinetics as the wild-type receptor. However, mutation of serine and/or threonine residues within the extreme carboxy-terminal attenuated internalization when these receptors were expressed in HEK293 cells. These results establish that the extreme carboxy-terminal phosphorylation sites are not required for internalization of truncated receptors, but are required for internalization of full-length receptors in HEK293 cells. Analysis of βarrestin-2 recruitment to mutant CB1 receptors suggests that putative carboxy-terminal phosphorylation sites mediate βarrestin-2 translocation. This study indicates that the local cellular environment affects the structural determinants of CB1 receptor internalization. Additionally, phosphorylation likely regulates the internalization of (full-length) CB1 receptors.
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影响因子：
2.9
作者：
Keren, O;Sarne, Y
通讯作者：
Sarne, Y
影响因子：
4.7
作者：
Hsieh, C;Brown, S;Mackie, K
通讯作者：
Mackie, K
影响因子：
5
作者：
Breivogel, CS;Scates, SM;Martin, BR
通讯作者：
Martin, BR
影响因子：
56.9
作者：
Berghuis, Paul;Rajnicek, Ann M.;Harkany, Tibor
通讯作者：
Harkany, Tibor
高渗培养基通过阻断网状蛋白包被的凹坑的形成来抑制受体介导的内吞作用。
DOI：
10.1083/jcb.108.2.389
发表时间：
1989-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Heuser JE;Anderson RG
通讯作者：
Anderson RG