Chemical proteomics tracks virus entry and uncovers NCAM1 as Zika virus receptor

• Published in: Nature communications Vol. 11; no. 1; pp. 3896 - 10
• Main Authors: Srivastava, Mayank, Zhang, Ying, Chen, Jian, Sirohi, Devika, Miller, Andrew, Zhang, Yang, Chen, Zhilu, Lu, Haojie, Xu, Jianqing, Kuhn, Richard J., Andy Tao, W.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 04.08.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 631/326/596/2557; 631/45/475; 631/92/96; 82/58; 96; Animals; Biotin; CD56 Antigen - genetics; CD56 Antigen - metabolism; Cell adhesion; Cell adhesion & migration; Cell adhesion molecules; Cell Line, Tumor; Chlorocebus aethiops; Gene Knockout Techniques; Glioblastoma; Glioblastoma cells; HEK293 Cells; Host-Pathogen Interactions - physiology; Humanities and Social Sciences; Humans; Internalization; multidisciplinary; Neural cell adhesion molecule; Neural Cell Adhesion Molecules - genetics; Neural Cell Adhesion Molecules - metabolism; Proteins; Proteomics; Receptors; Receptors, Virus - metabolism; Science; Science (multidisciplinary); Spectrometry; Tracks (paths); Ultraviolet radiation; Vector-borne diseases; Vero Cells; Viral Proteins - metabolism; Virus Attachment; Virus Internalization; Virus Replication - physiology; Viruses; Zika virus; Zika Virus - physiology; Zika Virus Infection - virology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-17638-y

The outbreak of Zika virus (ZIKV) in 2016 created worldwide health emergency which demand urgent research efforts on understanding the virus biology and developing therapeutic strategies. Here, we present a time-resolved chemical proteomic strategy to track the early-stage entry of ZIKV into host cells. ZIKV was labeled on its surface with a chemical probe, which carries a photocrosslinker to covalently link virus-interacting proteins in living cells on UV exposure at different time points, and a biotin tag for subsequent enrichment and mass spectrometric identification of the receptor or other host proteins critical for virus internalization. We identified Neural Cell Adhesion Molecule (NCAM1) as a potential ZIKV receptor and further validated it through overexpression, knockout, and inhibition of NCAM1 in Vero cells and human glioblastoma cells U-251 MG. Collectively, the strategy can serve as a universal tool to map virus entry pathways and uncover key interacting proteins. The mechanism underlying the cellular entry of Zika virus is not fully understood. Here, the authors use a chemically modified virus and time-resolved proteomics to capture interacting host proteins during virus entry and identify NCAM1 as a ZIKV receptor.