Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and (19)F-NMR.
Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and (19)F-NMR.
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非天然氨基酸掺入和 (19)F-NMR 揭示了视紫红质抑制蛋白构象和功能的磷酸选择性机制。
DOI：
10.1038/ncomms9202
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发表时间：
2015-09-08
影响因子：
16.6
通讯作者：
Sun JP
中科院分区：
文献类型：
--
作者：
Yang F;Yu X;Liu C;Qu CX;Gong Z;Liu HD;Li FH;Wang HM;He DF;Yi F;Song C;Tian CL;Xiao KH;Wang JY;Sun JP
特定的阻滞蛋白构象与不同的下游效应物偶联，这是许多g蛋白偶联受体（gpcr）功能的基础。在这里，使用非天然氨基酸掺入和氟-19核磁共振（19F-NMR）波谱，我们证明了不同的受体磷酸条形码被翻译成特定的β-arrestin-1构象和直接选择性信号传导。β-arrestin-1具有与磷酸结合的凹表面，可以“读取”受体磷酸- c尾部的信息，并通过19F-NMR揭示了不同的磷酸相互作用模式。虽然所有功能性磷酸肽都与一个共同的磷酸结合位点相互作用，并诱导手指环和中间环的运动，但不同的磷酸相互作用模式诱导不同的β-阻滞蛋白1的结构状态，这些结构状态与不同的阻滞蛋白功能相耦合。只有网格蛋白识别和稳定grk2特异性β-阻滞蛋白1构象。已确定的阻滞蛋白构象的受体-磷酸化选择机制以及阻滞蛋白中多个磷酸盐结合位点的间隔使阻滞蛋白能够识别许多gpcr的过度磷酸化状态，从而促进受体的功能多样性。G蛋白偶联受体（gpcr）通过G蛋白或阻滞蛋白介导的途径发出信号；抑制蛋白的可塑性被认为是其功能的基础。在这里，作者使用核磁共振来研究β-arrestin-1如何识别不同的GPCR磷酸条形码，以及这如何触发结构重排以实现选择性功能。
Specific arrestin conformations are coupled to distinct downstream effectors, which underlie the functions of many G-protein-coupled receptors (GPCRs). Here, using unnatural amino acid incorporation and fluorine-19 nuclear magnetic resonance (19F-NMR) spectroscopy, we demonstrate that distinct receptor phospho-barcodes are translated to specific β-arrestin-1 conformations and direct selective signalling. With its phosphate-binding concave surface, β-arrestin-1 ‘reads' the message in the receptor phospho-C-tails and distinct phospho-interaction patterns are revealed by 19F-NMR. Whereas all functional phosphopeptides interact with a common phosphate binding site and induce the movements of finger and middle loops, different phospho-interaction patterns induce distinct structural states of β-arrestin-1 that are coupled to distinct arrestin functions. Only clathrin recognizes and stabilizes GRK2-specific β-arrestin-1 conformations. The identified receptor-phospho-selective mechanism for arrestin conformation and the spacing of the multiple phosphate-binding sites in the arrestin enable arrestin to recognize plethora phosphorylation states of numerous GPCRs, contributing to the functional diversity of receptors. G-protein-coupled receptors (GPCRs) signal via G proteins or arrestin-mediated pathways; the plasticity of arrestin proteins is thought to underlie their function. Here, the authors use NMR to examine how β-arrestin-1 recognizes different GPCR phospho-barcodes, and how this triggers structural rearrangements to fulfill selective functions.