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 - Funding Information:
This work was supported in part by the NIH/NCI IMAT R33-CA186790-01A1 (to H.Z. and P.J.D.), NIH TCNP Roadmap (to H.Z.), NIH R01AG061852 (to H.Z.), NIH NS091102, AI84984, AI113273, and AI126176 (to K.S.K), NIH R01NS054791 (to X.D.), Johns Hopkins University Discovery Awards (to J.X.), Hamilton Innovation Award (to J. X.), Taiwanese Government Scholarship Award (to S.-C.W.), CDI Laboratories, and MOST Postdoctoral Fellowship—Taiwan (to G.-D.S.). We would like to thank the following colleagues for their assistance and support: David Leib (Dartmouth University) for the gift of KOS-37 BAC; David Johnson at Oregon Health and Science University for the generous gift of anti-gD antibody, DL6. We also thank Jen Tullman and Wade Gibson (Johns Hopkins University) for helpful discussions on virion purification and BAC recombineering method.
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.
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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 -