An Unbiased Screen for Human Cytomegalovirus Identifies Neuropilin-2 as a Central Viral Receptor

Characterizing cell surface receptors mediating viral infection is critical for understanding viral tropism and developing antiviral therapies. Nevertheless, due to challenges associated with detecting protein interactions on the cell surface, the host receptors of many human pathogens remain unknown. Here, we build a library consisting of most single transmembrane human receptors and implement a workflow for unbiased and high-sensitivity detection of receptor-ligand interactions. We apply this technology to elucidate the long-sought receptor of human cytomegalovirus (HCMV), the leading viral cause of congenital birth defects. We identify neuropilin-2 (Nrp2) as the receptor for HCMV-pentamer infection in epithelial/endothelial cells and uncover additional HCMV interactors. Using a combination of biochemistry, cell-based assays, and electron microscopy, we characterize the pentamer-Nrp2 interaction and determine the architecture of the pentamer-Nrp2 complex. This work represents an important approach to the study of host-pathogen interactions and provides a framework for understanding HCMV infection, neutralization, and the development of novel anti-HCMV therapies. [Display omitted] •High-throughput screen for unbiased detection of low-affinity pathogen-host interaction•Identification of novel interactors for HCMV entry complexes•Nrp2 is the HCMV pentamer receptor in epithelial and endothelial cells•Pentamer-specific HCMV neutralizing antibodies block Nrp2 binding A high-throughput screen for high-sensitivity detection of receptor-ligand interactions is used to identify long-sought cell-surface receptors responsible for human cytomegalovirus (HCMV) infection.