The rhodopsin-arrestin-1 interaction in bicelles.

The rhodopsin-arrestin-1 interaction in bicelles.
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Bicelles 中的视紫红质 - 抑制蛋白 - 1 相互作用。

DOI:
10.1007/978-1-4939-2330-4_6
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发表时间:
2015
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Iverson,TM
Iverson,TM
中科院分区:
--
文献类型:
--
作者:
Chen,Qiuyan;Vishnivetskiy,SergeyA;Zhuang,Tiandi;Cho,Min-Kyu;Thaker,TarjaniM;Sanders,CharlesR;Gurevich,VsevolodV;Iverson,TM

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G蛋白偶联受体(GPCRs)是真核细胞中信息传递的重要介质。GPCR与其结合伴侣之间的相互作用调节信号传导过程。例如,GPCR和同源G蛋白之间的相互作用启动信号,而与同源抑制蛋白的相互作用终止G蛋白介导的信号传导。在视觉信号转导中,arrestin-1选择性地与磷酸化的光激活GPCR视紫红质结合以终止视紫红质信号转导。在生理条件下,视紫红质-arrestin-1相互作用发生在视紫红质所在的高度特化的盘膜中。当与纯化的蛋白质一起工作时,该膜被模拟物取代。虽然洗涤剂通常用作膜模拟物,但大多数洗涤剂使arrestin-1变性,从而阻止了这种相互作用的生物化学研究。相反,双胞提供了一种合适的替代介质。双胞的一个优点是它们含有脂质,已经证明脂质是正常视紫红质-抑制蛋白-1相互作用所必需的。在这里,我们描述了如何将视紫红质重建为双胞,以及双胞性质如何影响视紫红质-抑制蛋白-1相互作用。
G-protein-coupled receptors (GPCRs) are essential mediators of information transfer in eukaryotic cells. Interactions between GPCRs and their binding partners modulate the signaling process. For example, the interaction between GPCR and cognate G protein initiates the signal, while the interaction with cognate arrestin terminates G-protein-mediated signaling. In visual signal transduction, arrestin-1 selectively binds to the phosphorylated light-activated GPCR rhodopsin to terminate rhodopsin signaling. Under physiological conditions, the rhodopsin-arrestin-1 interaction occurs in highly specialized disk membrane in which rhodopsin resides. This membrane is replaced with mimetics when working with purified proteins. While detergents are commonly used as membrane mimetics, most detergents denature arrestin-1, preventing biochemical studies of this interaction. In contrast, bicelles provide a suitable alternative medium. An advantage of bicelles is that they contain lipids, which have been shown to be necessary for normal rhodopsin-arrestin-1 interaction. Here we describe how to reconstitute rhodopsin into bicelles, and how bicelle properties affect the rhodopsin-arrestin-1 interaction.
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