Electrostatically-driven fast association and perdeuteration allow detection of transferred cross-relaxation for G protein-coupled receptor ligands with equilibrium dissociation constants in the high-to-low nanomolar range
Electrostatically-driven fast association and perdeuteration allow detection of transferred cross-relaxation for G protein-coupled receptor ligands with equilibrium dissociation constants in the high-to-low nanomolar range
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DOI：
10.1007/s10858-011-9523-3
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发表时间：
2011-07-01
影响因子：
2.7
通讯作者：
Baneres, Jean-Louis
中科院分区：
文献类型：
--
作者：
Catoire, Laurent J.;Damian, Marjorie;Baneres, Jean-Louis
G蛋白偶联受体介导的信号转导机制是药理学和结构生物学研究的热点。配体与受体的结合构成了激活过程中的关键事件。即使平衡解离常数低于微摩尔范围，溶液状态核磁共振也可以通过检测转移模式下的偶极相互作用来高分辨率确定受体结合状态下的激动剂分子的结构。这是可能的，在这种情况下，带电配体固有的超快扩散结合到高度带电的细胞外表面，并通过过氢化受体来减缓H-1-H-1交叉松弛。在这里，我们为两个与白三烯BLT2受体相互作用的脂肪酸分子证明了这一点，这两个配体都表现出亚微摩尔亲和力。
The mechanism of signal transduction mediated by G protein-coupled receptors is a subject of intense research in pharmacological and structural biology. Ligand association to the receptor constitutes a critical event in the activation process. Solution-state NMR can be amenable to high-resolution structure determination of agonist molecules in their receptor-bound state by detecting dipolar interactions in a transferred mode, even with equilibrium dissociation constants below the micromolar range. This is possible in the case of an inherent ultra-fast diffusive association of charged ligands onto a highly charged extracellular surface, and by slowing down the H-1-H-1 cross-relaxation by perdeuterating the receptor. Here, we demonstrate this for two fatty acid molecules in interaction with the leukotriene BLT2 receptor, for which both ligands display a submicromolar affinity.