TY - JOUR
T1 - Development and application of a high-content virion display human GPCR array
AU - Syu, Guan Da
AU - Wang, Shih Chin
AU - Ma, Guangzhong
AU - Liu, Shuang
AU - Pearce, Donna
AU - Prakash, Atish
AU - Henson, Brandon
AU - Weng, Lien Chun
AU - Ghosh, Devlina
AU - Ramos, Pedro
AU - Eichinger, Daniel
AU - Pino, Ignacio
AU - Dong, Xinzhong
AU - Xiao, Jie
AU - Wang, Shaopeng
AU - Tao, Nongjian
AU - Kim, Kwang Sik
AU - Desai, Prashant J.
AU - Zhu, Heng
N1 - Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85064942092&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85064942092&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-09938-9
DO - 10.1038/s41467-019-09938-9
M3 - Article
C2 - 31040288
AN - SCOPUS:85064942092
SN - 2041-1723
VL - 10
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 1997
ER -