A Suitable Membrane Distance Regulated by the RBD_ACE2 Interaction is Critical for SARS-CoV-2 Spike-Mediated Viral Invasion
© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH..
The receptor-binding domain (RBD) of spike recognizing the receptor angiotensin-converting enzyme 2 (ACE2) initiates membrane fusion between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and cell membrane. Although the structure of the RBD_ACE2 complex has been well studied, its functional mechanism in membrane fusion is still not fully understood. Here, using an in vitro cell-vesicle content-mixing assay, it is found that the cleavage at the S2' site by thrombin (Thr) protease strongly accelerates membrane fusion, compared to that of cleavage at the S1/S2 site by PreScission (3C) protease. Moreover, mutations at the RBD_ACE2 interface resulted in a positive correlation between binding affinity and fusion probability. In both the cell-vesicle and cell-cell fusion assays, by crosslinking two membranes via the neutravidin (NTV)_biotin interaction or complementary DNA strands, it is found that spike drives membrane fusion in the absence of ACE2, and a suitable distance between two membranes is critical for spike-mediated membrane fusion. Finally, unsuitable membrane crosslinkers significantly inhibited the fusion probability in the presence of ACE2. Taken together, the results suggest that the RBD_ACE2 complex may act as a crosslinker to bridge the viral and cell membranes at a suitable distance, which is critical, but also substitutable for spike-mediated SARS-CoV-2 entry.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:10 |
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| Enthalten in: |
Advanced science (Weinheim, Baden-Wurttemberg, Germany) - 10(2023), 28 vom: 17. Okt., Seite e2301478 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wu, Mengdan [VerfasserIn] |
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| Links: |
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| Themen: |
Journal Article |
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| Anmerkungen: |
Date Revised 10.10.2023 published: Print-Electronic Citation Status Publisher |
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| doi: |
10.1002/advs.202301478 |
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| 520 | |a The receptor-binding domain (RBD) of spike recognizing the receptor angiotensin-converting enzyme 2 (ACE2) initiates membrane fusion between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and cell membrane. Although the structure of the RBD_ACE2 complex has been well studied, its functional mechanism in membrane fusion is still not fully understood. Here, using an in vitro cell-vesicle content-mixing assay, it is found that the cleavage at the S2' site by thrombin (Thr) protease strongly accelerates membrane fusion, compared to that of cleavage at the S1/S2 site by PreScission (3C) protease. Moreover, mutations at the RBD_ACE2 interface resulted in a positive correlation between binding affinity and fusion probability. In both the cell-vesicle and cell-cell fusion assays, by crosslinking two membranes via the neutravidin (NTV)_biotin interaction or complementary DNA strands, it is found that spike drives membrane fusion in the absence of ACE2, and a suitable distance between two membranes is critical for spike-mediated membrane fusion. Finally, unsuitable membrane crosslinkers significantly inhibited the fusion probability in the presence of ACE2. Taken together, the results suggest that the RBD_ACE2 complex may act as a crosslinker to bridge the viral and cell membranes at a suitable distance, which is critical, but also substitutable for spike-mediated SARS-CoV-2 entry | ||
| 650 | 4 | |a Journal Article | |
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| 650 | 4 | |a SARS-CoV-2 | |
| 650 | 4 | |a membrane fusion | |
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| 700 | 1 | |a Fan, Jiaqi |e verfasserin |4 aut | |
| 700 | 1 | |a Cui, Lele |e verfasserin |4 aut | |
| 700 | 1 | |a Xiang, Yijuan |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Kaiyu |e verfasserin |4 aut | |
| 700 | 1 | |a Tang, Linwei |e verfasserin |4 aut | |
| 700 | 1 | |a Duan, Yanping |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Zimin |e verfasserin |4 aut | |
| 700 | 1 | |a Yang, Fanli |e verfasserin |4 aut | |
| 700 | 1 | |a Shui, Weiwei |e verfasserin |4 aut | |
| 700 | 1 | |a Lu, Guangwen |e verfasserin |4 aut | |
| 700 | 1 | |a Lai, Ying |e verfasserin |4 aut | |
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