Differential manipulation of arrestin-3 binding to basal and agonist-activated G protein-coupled receptors

作者：Prokop Susanne; Perry Nicole A; Vishnivetskiy Sergey A; Toth Andras D; Inoue Asuka; Milligan Graeme; Iverson Tina M; Hunyady Laszlo; Gurevich Vsevolod V^*

来源：Cellular Signalling, 2017, 36: 98-107.

DOI：10.1016/j.cellsig.2017.04.021

• Arrestin
• GPCRs
• Receptor specificity
• Protein-protein interactions
• Protein engineering

Non-visual arrestins interact with hundreds of different G protein-coupled receptors (GPCRs). Here we show that by introducing mutations into elements that directly bind receptors, the specificity of arrestin-3 can be altered. Several mutations in the two parts of the central "crest" of the arrestin molecule, middle-loop and C-loop, enhanced or reduced arrestin-3 interactions with several GPCRs in receptor subtype and functional state-specific manner. For example, the Lys139Ile substitution in the middle-loop dramatically enhanced the binding to inactive M-2 muscarinic receptor, so that agonist activation of the M-2 did not further increase arrestin-3 binding. Thus, the Lys139Ile mutation made arrestin-3 essentially an activation-independent binding partner of M-2, whereas its interactions with other receptors, including the 112-adrenergic receptor and the D-1 and D-2 dopamine receptors, retained normal activation dependence. In contrast, the Ala248Val mutation enhanced agonistinduced arrestin-3 binding to the beta(2)-adrenergic and D-2 dopamine receptors, while reducing its interaction with the D-1 dopamine receptor. These mutations represent the first example of altering arrestin specificity via enhancement of the arrestin-receptor interactions rather than selective reduction of the binding to certain subtypes.

• 出版日期2017-8