Plasmon-waveguide resonance studies of ligand binding to the human β2-adrenergic receptor
Plasmon-waveguide resonance studies of ligand binding to the human β2-adrenergic receptor
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DOI：
10.1021/bi035825a
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发表时间：
2004-03-23
期刊：
影响因子：
2.9
通讯作者：
Tollin, G
中科院分区：
文献类型：
--
作者：
Devanathan, S;Yao, ZP;Tollin, G
等离子体波导共振（PWR）光谱是一种光学技术，可用于实时探测各向异性蛋白脂膜内发生的分子相互作用，而无需进行分子标记。该方法直接监测在此类系统中发生的质量密度、构象和分子取向变化，并允许测定蛋白质-配体结合常数和结合动力学。在目前的研究中，PWR已被用于监测人β(2)-肾上腺素能受体与固体支撑的鸡蛋磷脂酰胆碱脂双分子层的结合，并跟踪完全激动剂（异丙肾上腺素、肾上腺素）、部分激动剂（多巴酚丁胺）、拮抗剂（阿普利诺尔）和逆激动剂（ci -118,551）与受体的结合。结合动力学的差异和PWR光谱的变化表明存在多种配体类型特征的构象，反映了结合过程中产生的受体结构状态的差异。这些结果为β(2)-肾上腺素能受体形成的配位态的构象异质性提供了新的证据。
Plasmon-waveguide resonance (PWR) spectroscopy is an optical technique that can be used to probe the molecular interactions occurring within anisotropic proteolipid membranes in real time without requiring molecular labeling. This method directly monitors mass density, conformation, and molecular orientation changes occurring in such systems and allows determination of protein-ligand binding constants and binding kinetics. In the present study, PWR has been used to monitor the incorporation of the human beta(2)-adrenergic receptor into a solid-supported egg phosphatidylcholine lipid bilayer and to follow the binding of full agonists (isoproterenol, epinephrine), a partial agonist (dobutamine), an antagonist (alprenolol), and an inverse agonist (ICI-118,551) to the receptor. The combination of differences in binding kinetics and the PWR spectral changes point to the occurrence of multiple conformations that are characteristic of the type of ligand, reflecting differences in the receptor structural states produced by the binding process. These results provide new evidence for the conformational heterogeneity of the liganded states formed by the beta(2)-adrenergic receptor.