Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

Ke Wang; Yu Pan; Dianbing Wang; Ye Yuan; Min Li; Yuanyuan Chen; Lijun Bi; Xian-En Zhang

doi:10.1016/j.virs.2023.06.005

August 2023

. doi: 10.1016/j.virs.2023.06.005

Citation: Ke Wang, Yu Pan, Dianbing Wang, Ye Yuan, Min Li, Yuanyuan Chen, Lijun Bi, Xian-En Zhang. Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry .VIROLOGICA SINICA, 2023, 38(4) : 595-605. http://dx.doi.org/10.1016/j.virs.2023.06.005

SARS-CoV-2刺突蛋白突变体与hACE2结合亲和力及S1/S2切割效率的改变影响病毒进入细胞

王坷 ^a,b,c ,
潘瑜 ^a ,
王殿冰 ^a ,
袁也 ^a ,
李敏 ^a ,
陈媛媛 ^a ,
毕利军 ^a ,
张先恩 ^a,b,c,,

中国科学院生物物理研究所;中国科学院深圳先进技术研究院;中国科学院大学

通讯作者： 张先恩, zhangxe@ibp.ac.cn
收稿日期： 2023-01-31
录用日期： 2023-06-15

摘要

新型冠状病毒变异株不断出现，影响了公共卫生措施对感染人数的控制。虽然已经观察到不同变异株的刺突蛋白突变直接影响病毒进入宿主细胞，但仍需要进一步对这些变异体进行系统、多方面的比较。本研究系统研究了八种关注变异株（VOC）和感兴趣变异株（VOI）的刺突蛋白突变对蛋白表达量、蛋白酶切割效率、与受体的结合亲和力、病毒进入细胞效率及宿主细胞嗜性的影响。发现Omicron的全长刺突蛋白和其受体结合结构域（RBD）的亲和力与野生型的相似。此外，Alpha、Beta、Delta和Lambda假病毒相比野生型获得了显著增强的进入细胞能力，Omicron假病毒进入细胞的能力略有增加，这表明Omicron变异株显著增加的传播速度与其刺突蛋白和hACE2之间的亲和力无关。还发现Omicron和Mu的刺突蛋白在S1/S2位点被切割的效率较低，且不能有效利用TMPRSS2进入宿主细胞，表明它们更倾向于通过内吞作用途径进入宿主细胞。此外，我们测试的所有变异株假病毒都获得了进入表达动物ACE2细胞的能力，尤其是对大鼠和小鼠的感染潜力显著增加，这表示啮齿动物可能成为病毒进化的储备库。本研究的发现为针对性的流行病控制提供了有价值的指导，有助于更好地理解SARS-CoV-2的演化。
- 新型冠状病毒2型
- , 关注变异株（VOC）
- , 奥密克戎
- , 结合亲和力
- , 病毒进入
- , 宿主嗜性

Altered hACE2 binding affinity and S1/S2 cleavage efficiency of SARS-CoV-2 spike protein mutants affect viral cell entry

Ke Wang ^a,b,c ,
Yu Pan ^a ,
Dianbing Wang ^a ,
Ye Yuan ^a ,
Min Li ^a ,
Yuanyuan Chen ^a ,
Lijun Bi ^a ,
Xian-En Zhang ^a,b,c,,

a. National Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China;
b. Faculty of Synthetic Biology, Shenzhen Institute of Advanced Technology, Shenzhen, 518055, China;
c. University of Chinese Academy of Sciences, Beijing, 100101, China

Corresponding author: Xian-En Zhang, zhangxe@ibp.ac.cn
Received Date: 31 January 2023
Accepted Date: 15 June 2023

Abstract

SARS-CoV-2 variants are constantly emerging, hampering public health measures in controlling the number of infections. While it is well established that mutations in spike proteins observed for the different variants directly affect virus entry into host cells, there remains a need for further expansion of systematic and multifaceted comparisons. Here, we comprehensively studied the effect of spike protein mutations on spike expression and proteolytic activation, binding affinity, viral entry efficiency and host cell tropism of eight variants of concern (VOC) and variants of interest (VOI). We found that both the full-length spike and its receptor-binding domain (RBD) of Omicron bind to hACE2 with an affinity similar to that of the wild-type. In addition, Alpha, Beta, Delta and Lambda pseudoviruses gained significantly enhanced cell entry ability compared to the wild-type, while the Omicron pseudoviruses showed a slightly increased cell entry, suggesting the vastly increased rate of transmission observed for Omicron variant is not associated with its affinity to hACE2. We also found that the spikes of Omicron and Mu showed lower S1/S2 cleavage efficiency and inefficiently utilized TMPRSS2 to enter host cells than others, suggesting that they prefer the endocytosis pathway to enter host cells. Furthermore, all variants' pseudoviruses we tested gained the ability to enter the animal ACE2-expressing cells. Especially the infection potential of rats and mice showed significantly increased, strongly suggesting that rodents possibly become a reservoir for viral evolution. The insights gained from this study provide valuable guidance for a targeted approach to epidemic control, and contribute to a better understanding of SARS-CoV-2 evolution.
- SARS-CoV-2
- , Variants of Concern (VOC)
- , Omicron
- , Binding affinity
- , Viral entry
- , Host-tropism

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10.1016j.virs.2023.06.005-ESM.doc