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Nat Commun. 2019 Apr 30;10(1):1997. doi: 10.1038/s41467-019-09938-9.

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

Author information

1
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
2
Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
3
Viral Oncology Program, Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21231, USA.
4
Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
5
Biodesign Center for Bioelectronics and Biosensors, Arizona State University, Tempe, AZ, 85287, USA.
6
Division of Paediatric Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA.
7
Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
8
CDI Laboratories, Inc., Mayaguez, Puerto Rico, 00682, USA.
9
Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.
10
School of Electrical, Computer and Energy Engineering, Arizona State University, Tempe, AZ, 85287, USA.
11
Division of Paediatric Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA. kwangkim@jhmi.edu.
12
Viral Oncology Program, Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21231, USA. pdesai@jhmi.edu.
13
Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA. hzhu4@jhmi.edu.
14
Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA. hzhu4@jhmi.edu.
15
Viral Oncology Program, Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21231, USA. hzhu4@jhmi.edu.

Abstract

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.

PMID:
31040288
PMCID:
PMC6491619
DOI:
10.1038/s41467-019-09938-9
[Indexed for MEDLINE]
Free PMC Article

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