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Development and application of a high-content virion display human GPCR array.

Guan-Da SyuShih-Chin WangGuangzhong MaShuang LiuDonna PearceAtish PrakashBrandon HensonLien-Chun WengDevlina GhoshPedro RamosDaniel EichingerIgnacio PinoXinzhong DongJie XiaoShaopeng WangNongjian TaoKwang Sik KimPrashant J DesaiHeng Zhu
Published in: Nature communications (2019)
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.
Keyphrases
  • high throughput
  • high resolution
  • endothelial cells
  • small molecule
  • high density
  • induced pluripotent stem cells
  • pluripotent stem cells
  • risk assessment
  • mass spectrometry
  • pseudomonas aeruginosa
  • drug induced