The interplay of receptor dynamics and ligand residence time upon ligand efficacy at adenosine receptors (C02)

导致配体停留时间增加的分子细节目前还不充分了解。这个项目的主要问题将是在活细胞中使用高动力学分辨率的fret显微镜来解决与配体结合有关的受体构象动力学。利用腺苷受体，我们将：1)研究细胞外环2在配体结合动力学中的作用；II)比较低配体亲和力的A2B受体与高亲和力的A1和A2A受体；III)研究动态配体结合与A1受体上H251A突变构象动力学的关系；IV)研究g蛋白对受体动态和受体定位的影响；V)以PD 81723为原型，研究变构调制剂在停留时间和信号传导中的作用；VI)使系统适应96孔板，以提高吞吐量。

The molecular details leading to increased ligand residence time are currently insufficiently understood. The major question within this project will be to address conformational dynamic of a receptor in relation to ligand binding using FRET-microscopy with high kinetic resolution in living cells. Using adenosine receptors, we will: I) Study the role of the extracellular loop 2 in ligand binding kinetics; II) Compare the low ligand affinity A2B receptor to the high affinity A1 and A2A receptors; III) Investigate dynamic ligand binding in relation to the conformational dynamics of the H251A mutation at the A1 receptor; IV) Study the influence of G-proteins in receptor dynamics and receptor localization; V) Investigate the role of allosteric modulators in residence time and signaling using PD 81,723 as prototype; VI) Adapt the system to 96-well plates for faster throughput.