Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
Nat Commun. 2019 Apr 30;10(1):1997. doi: 10.1038/s41467-019-09938-9.
Human G protein-coupled receptors (GPCRs) respond to various ligands and stimuli. However, GPCRs rely on membrane for proper folding, making their biochemical properties difficult to study. By displaying GPCRs in viral envelopes, we fabricated a Virion Display (VirD) array containing 315 non-olfactory human GPCRs for functional characterization. Using this array, we found that 10 of 20 anti-GPCR mAbs were ultra-specific. We further demonstrated that those failed in the mAb assays could recognize their canonical ligands, suggesting proper folding. Next, using two peptide ligands on the VirD-GPCR array, we identified expected interactions and novel interactions. Finally, we screened the array with group B Streptococcus, a major cause of neonatal meningitis, and demonstrated that inhibition of a newly identified target, CysLTR1, reduced bacterial penetration both in vitro and in vivo. We believe that the VirD-GPCR array holds great potential for high-throughput screening for small molecule drugs, affinity reagents, and ligand deorphanization.
人 G 蛋白偶联受体 (GPCRs) 可响应各种配体和刺激。然而,GPCRs 的正确折叠依赖于膜,这使得它们的生化特性难以研究。通过在病毒包膜中展示 GPCRs,我们构建了一个包含 315 种非嗅觉人类 GPCR 的病毒展示 (VirD) 阵列,用于功能表征。使用该阵列,我们发现 20 种抗 GPCR mAb 中有 10 种具有超高特异性。我们进一步证明,那些在 mAb 测定中失败的 mAb 可以识别它们的经典配体,表明其正确折叠。接下来,我们使用 VirD-GPCR 阵列上的两个肽配体,鉴定了预期的和新的相互作用。最后,我们用 B 群链球菌(新生儿脑膜炎的主要病因)对该阵列进行了筛选,并证明抑制新鉴定的靶点 CysLTR1 可减少细菌在体外和体内的穿透。我们相信,VirD-GPCR 阵列在高通量筛选小分子药物、亲和试剂和配体鉴定方面具有巨大潜力。