Structural and functional constraints on spike activation and host protease utilization limit cell entry of SARS-CoV-2-related bat coronaviruses
The viral entry mechanisms, which are primarily related to the spike character, play a critical role in determining zoonotic potential. Among the currently identified SC2r-CoVs, BANAL-52 and BANAL-103 exhibit spike proteins with the highest sequence similarity to SARS-CoV-2, rendering them optimal models for comparative studies on S-mediated cell entry and cross-species transmission. In this study, we systematically investigated the molecular constraints governing the functionality of BANAL spikes, with a focus on S-ACE2 interactions, S activation, S structures, and host protease utilization. Notably, we resolved the cryo-EM structure of BANAL-52 S at neutral pH and the first cryo-EM structure of BANAL-103 S, revealing distinct glycan- and lipid-mediated stabilization of inactive states. Furthermore, cross-neutralization assays demonstrated that sera of convalescents from SARS-CoV-2 inhibited BANAL pseudovirus entry with an efficiency of approximately 80%, thereby highlighting conserved antigenic epitopes and informing the development of broad-spectrum therapeutic strategies against emerging SC2r-CoVs.
Authors
- Shuiling Jin (ORCID: https://orcid.org/0000-0001-7330-7140)
- Na Zang (ORCID: https://orcid.org/0000-0001-6658-6700)
- Jiaxu Wang (ORCID: https://orcid.org/0000-0001-7101-8230)
- Didi Wan
- Yi-Bing Zhang (ORCID: https://orcid.org/0000-0001-7647-9335)
- Xiangdong Sun (ORCID: https://orcid.org/0000-0002-0051-1463)
- Xiao-Qing Cai (ORCID: https://orcid.org/0000-0002-6025-2491)
- Jianxiang Shi (ORCID: https://orcid.org/0000-0002-4346-3895)
- Yushun Wan (ORCID: https://orcid.org/0000-0003-0496-6315)
- Jian Shang
- Zhengkun Xie (ORCID: https://orcid.org/0000-0001-7422-9527)
- Qingqing Li (ORCID: https://orcid.org/0000-0002-3166-3195)
- Junxia Yang
- Xiaoning Li (ORCID: https://orcid.org/0000-0002-1608-6887)
- Ru Li (ORCID: https://orcid.org/0000-0002-6275-5156)
- Zhijun Kang (ORCID: https://orcid.org/0009-0003-4234-6659)
- Lili Li (ORCID: https://orcid.org/0000-0003-0552-7250)
- Ying Peng (ORCID: https://orcid.org/0009-0002-2907-7741)
Institutions
- Zhengzhou University (CN)
- Fifth Affiliated Hospital of Zhengzhou University (CN)
- First Affiliated Hospital of Zhengzhou University (CN)
- Children's Hospital of Chongqing Medical University (CN)
- Henan Normal University (CN)
- Chongqing Medical University (CN)
Publication Details
- Journal
- Journal of Virology
- Published
- 2025-07-24
- DOI
- https://doi.org/10.1128/jvi.01007-25
- Citations
- 1
- Primary Topic
- SARS-CoV-2 and COVID-19 Research
- Type
- article
- Field-Weighted Citation Impact
- 1.40
Funders
- National Natural Science Foundation of China