Actin cytoskeleton-dependent dynamics of the human serotonin1A receptor correlates with receptor signaling
Actin cytoskeleton-dependent dynamics of the human serotonin1A receptor correlates with receptor signaling
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DOI：
10.1529/biophysj.107.125732
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发表时间：
2008-07-01
影响因子：
3.4
通讯作者：
Chattopadhyay, Amitabha
中科院分区：
文献类型：
--
作者：
Ganguly, Sourav;Pucadyil, Thomas J.;Chattopadhyay, Amitabha
在细胞信号传导的背景下分析膜蛋白的动力学是当代细胞生物学中具有挑战性的问题。已知细胞膜中脂质和蛋白质的侧向扩散受细胞骨架的影响。在这项工作中，我们探讨了肌动蛋白细胞骨架上的5-羟色胺（1A）（5-HT 1A）受体，在CHO细胞中稳定表达的流动性的作用，及其在信号转导的影响。5-HT 1AR-EYFP的FRAP分析表明，由CD或毛喉素介导的cAMP水平升高诱导的肌动蛋白细胞骨架的不稳定导致受体的移动的部分增加。移动的分数的增加伴随着受体的信号传导效率的相应增加。有趣的是，随着使用的CD浓度的增加，移动的部分的增加与受体的配体介导的信号传导效率表现出类似于0.95的相关性。放射性配体结合和G-蛋白偶联的受体被认为是不受影响的CD治疗后。我们的研究结果表明，5-羟色胺（1A）受体的信号与受体的流动性，从而意味着肌动蛋白细胞骨架可以发挥调节作用的受体信号。这些结果可能具有潜在的意义的背景下，一般的GPCR信号，并在GPCR-细胞骨架相互作用的理解与受体信号，特别是。
Analyzing the dynamics of membrane proteins in the context of cellular signaling represents a challenging problem in contemporary cell biology. Lateral diffusion of lipids and proteins in the cell membrane is known to be influenced by the cytoskeleton. In this work, we explored the role of the actin cytoskeleton on the mobility of the serotonin(1A) (5-HT1A) receptor, stably expressed in CHO cells, and its implications in signaling. FRAP analysis of 5-HT1AR-EYFP shows that destabilization of the actin cytoskeleton induced by either CD or elevation of cAMP levels mediated by forskolin results in an increase in the mobile fraction of the receptor. The increase in the mobile fraction is accompanied by a corresponding increase in the signaling efficiency of the receptor. Interestingly, with increasing concentrations of CD used, the increase in the mobile fraction exhibited a correlation of similar to 0.95 with the efficiency in ligand-mediated signaling of the receptor. Radioligand binding and G-protein coupling of the receptor were found to be unaffected upon treatment with CD. Our results suggest that signaling by the serotonin(1A) receptor is correlated with receptor mobility, implying thereby that the actin cytoskeleton could play a regulatory role in receptor signaling. These results may have potential significance in the context of signaling by GPCRs in general and in the understanding of GPCR-cytoskeleton interactions with respect to receptor signaling in particular.