Nanobody-Enabled Reverse Pharmacology on G-Protein-Coupled Receptors
Nanobody-Enabled Reverse Pharmacology on G-Protein-Coupled Receptors
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DOI：
10.1002/anie.201712581
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发表时间：
2018-05-04
影响因子：
16.6
通讯作者：
Steyaert, Jan
中科院分区：
文献类型：
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作者：
Pardon, Els;Betti, Cecilia;Steyaert, Jan
g蛋白偶联受体（gpcr）跨膜信号的构象复杂性是设计受体激动剂筛选的中心障碍。在其基础状态下，与g蛋白结合的活性状态构象相比，gpcr对激动剂的亲和力较低。此外，不同的激动剂可以稳定不同的活性受体构象，并不能均匀地激活与给定受体相关的所有细胞信号通路（激动剂偏差）。比较片段筛选对β(2-)肾上腺素受体-纳米体融合被g蛋白模拟纳米体锁定在其活性状态构象和相同的受体在其基底状态构象。这种简单的生物物理分析方法可以对多种新型激动剂进行鉴定和排序，并对每种激动剂、拮抗剂或逆激动剂的功效进行分类，从而为纳米体激活的反向药理学打开了大门。
The conformational complexity of transmembrane signaling of G-protein-coupled receptors (GPCRs) is a central hurdle for the design of screens for receptor agonists. In their basal states, GPCRs have lower affinities for agonists compared to their G-protein-bound active state conformations. Moreover, different agonists can stabilize distinct active receptor conformations and do not uniformly activate all cellular signaling pathways linked to a given receptor (agonist bias). Comparative fragment screens were performed on a beta(2-)adrenoreceptor-nanobody fusion locked in its active-state conformation by a G-protein-mimicking nanobody, and the same receptor in its basal-state conformation. This simple biophysical assay allowed the identification and ranking of multiple novel agonists and permitted classification of the efficacy of each hit in agonist, antagonist, or inverse agonist categories, thereby opening doors to nanobody-enabled reverse pharmacology.