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Author Notes:

Wen‐xia Tian, Email: wenxiatian@126.com

WT, QW, and GFG initiated and coordinated the project. QW designed the experiments. SN and BB performed the SPR analysis, and SN. YJ, and QC conducted the flow cytometry assay. SN and AZ performed the pseudovirus infection assay. SN, BB, CQ, and YZ prepared the complexes of GX/P2V/2017 RBD‐hACE2 and GD/1/2019 RBD‐hACE2. JW and H‐WW prepared the samples and collected the structural data. JW and JQ solved the cryo‐EM structures. SN, LW, PH, QW, and GFG analyzed the data. SN, PD, QW, and GFG wrote the manuscript.

We are grateful to Zheng Fan (Institute of Microbiology, Chinese Academy of Sciences (CAS)) for their technical support of SPR analysis. We thank Jianlin Lei, Xiaomin Li at Tsinghua University for data collection. We thank the Tsinghua University Branch of the China National Center for Protein Sciences (Beijing) for providing the cryo‐EM facility support and the computational facility support on the cluster of Bio‐Computing Platform.

The authors declare that they have no conflict of interest.

Subjects:

Research Funding:

This work was supported by the Ministry of Science and Technology of the People’s Republic of China (2020YFC0840800 and 2020YFC0845900), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29010202), the intramural special grant for SARS‐CoV‐2 research from the Chinese Academy of Sciences, and the National Natural Science Foundation of China (81922044). Q.W. is supported by the Youth Innovation Promotion Association CAS (2018119). G.F.G is supported by the Yanqi Lake Meeting organized by the Academic Divisions of CAS.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemistry & Molecular Biology
  • Cell Biology
  • ACE2
  • COVID-19
  • Cryo-EM
  • pangolin CoVs
  • SARS-CoV-2
  • BROAD-HOST-RANGE
  • CORONAVIRUS
  • PATHOGENESIS
  • OUTBREAK
  • SPIKE
  • CHINA

Molecular basis of cross-species ACE2 interactions with SARS-CoV-2-like viruses of pangolin origin

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Journal Title:

EMBO JOURNAL

Volume:

Volume 40, Number 16

Publisher:

, Pages e107786-e107786

Type of Work:

Article | Final Publisher PDF

Abstract:

Pangolins have been suggested as potential reservoir of zoonotic viruses, including SARS-CoV-2 causing the global COVID-19 outbreak. Here, we study the binding of two SARS-CoV-2-like viruses isolated from pangolins, GX/P2V/2017 and GD/1/2019, to human angiotensin-converting enzyme 2 (hACE2), the receptor of SARS-CoV-2. We find that the spike protein receptor-binding domain (RBD) of pangolin CoVs binds to hACE2 as efficiently as the SARS-CoV-2 RBD in vitro. Furthermore, incorporation of pangolin CoV RBDs allows entry of pseudotyped VSV particles into hACE2-expressing cells. A screen for binding of pangolin CoV RBDs to ACE2 orthologs from various species suggests a broader host range than that of SARS-CoV-2. Additionally, cryo-EM structures of GX/P2V/2017 and GD/1/2019 RBDs in complex with hACE2 show their molecular binding in modes similar to SARS-CoV-2 RBD. Introducing the Q498H substitution found in pangolin CoVs into the SARS-CoV-2 RBD expands its binding capacity to ACE2 homologs of mouse, rat, and European hedgehog. These findings suggest that these two pangolin CoVs may infect humans, highlighting the necessity of further surveillance of pangolin CoVs.

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© 2021 The Authors

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/rdf).
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