Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2

Wang, Qihui; Zhang, Yanfang; Wu, Lili; Niu, Sheng; Song, Chunli; Zhang, Zengyuan; Lu, Guangwen; Qiao, Chengpeng; Hu, Yu; Yuen, Kwok Yung; Wang, Qisheng; Zhou, Huan; Yan, Jinghua; Qi, Jianxun

doi:10.1016/j.cell.2020.03.045

Cited by 2,801 publications

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“…Indeed, most of neutralizing monoclonal antibodies (mAbs) previously developed against SARS-CoV target the RBD epitopes, while a few are directed against the S2 subunit or the S1/S2 cleavage site ( 29, 30 ). The cross-reactivity of such mAbs with SARS-CoV-2 has been characterized, and it was found that many SARS-CoV-neutralizing mAbs exhibit no cross-neutralizing capacity ( 9, 31 ). For example, CR3022, a neutralizing antibody isolated from a convalescent SARS patient, cross-reacted with the RBD of SARS-CoV-2 but did not neutralize the virus ( 31, 32 ).…”

Section: Discussionmentioning

confidence: 99%

Cross-reactive neutralization of SARS-CoV-2 by serum antibodies from recovered SARS patients and immunized animals

Zhu

Han

et al. 2020

Preprint

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16The current COVID-19 pandemic, caused by a novel coronavirus SARS-CoV-2, poses serious 17 threats to public health and social stability, calling for urgent need for vaccines and 18 therapeutics. SARS-CoV-2 is genetically close to SARS-CoV, thus it is important to define 19 the between antigenic cross-reactivity and neutralization. In this study, we firstly analyzed 20 20 convalescent serum samples collected from SARS-CoV infected individuals during the 2003 21 SARS outbreak. All patient sera reacted strongly with the S1 subunit and receptor-binding 22 domain (RBD) of SARS-CoV, cross-reacted with the S ectodomain, S1, RBD, and S2 23 proteins of SARS-CoV-2, and neutralized both SARS-CoV and SARS-CoV-2 S 24 protein-driven infections. Multiple panels of antisera from mice and rabbits immunized with a 25 full-length S and RBD immunogens of SARS-CoV were also characterized, verifying the 26 cross-reactive neutralization against SARS-CoV-2. Interestingly, we found that a palm civet 27 SARS-CoV-derived RBD elicited more potent cross-neutralizing responses in immunized 28 animals than the RBD from a human SARS-CoV strain, informing a strategy to develop a 29 universe vaccine against emerging CoVs. 30 31 Summary 32 Serum antibodies from SARS-CoV infected patients and immunized animals cross-neutralize 33 SARS-CoV-2 suggests strategies for universe vaccines against emerging CoVs. acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is a new coronavirus (CoV) 37 genetically close to SARS-CoV emerged in 2002 (1-3). As of 20 April 2020, a total of 38 2,246,291 confirmed COVID-19 cases, including 152,707 deaths, have been reported from 39 213 countries or regions, and the numbers are growing rapidly (https://www.who.int). The 40 pandemic threatens to become one of the most difficult times faced by humans in modern 41 history. Unfortunately, even though 17 years passed, we have not developed effective 42 prophylactics and therapeutics in preparedness for the re-emergence of SARS or SARS-like 43 CoVs. A vaccine is urgently needed to prevent the human-to-human transmission of 44 SARS-CoV-2. 45Like SARS-CoV and many other CoVs, SARS-CoV-2 utilizes its surface spike (S) 46 glycoprotein to gain entry into host cells (4-6). Typically, the S protein forms a homotrimer 47 with each protomer consisting of S1 and S2 subunits. The N-terminal S1 subunit is 48 responsible for virus binding to the cellular receptor ACE2 through an internal 49 receptor-binding domain (RBD) that is capable of functional folding independently, whereas 50 the membrane-proximal S2 subunit mediates membrane fusion events. Very recently, the 51 prefusion structure of the SARS-CoV-2 S protein was determined by cryo-EM, which 52 revealed an overall similarity to that of SARS-CoV (4, 7); the crystal structure of the 53 SARS-CoV-2 RBD in complex with ACE2 was also determined by several independent 54 groups, and the residues or motifs critical for the higher-affinity RBD-ACE2 interaction were 55 identified (8-10).The S protein of CoVs is also a main target of ...

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Section: Discussionmentioning

confidence: 99%

Cross-reactive neutralization of SARS-CoV-2 by serum antibodies from recovered SARS patients and immunized animals

Zhu

Han

et al. 2020

Preprint

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“…It has been proposed by biolayer interferometry studies that the receptorbinding domains of SARS-CoV-1 and SARS-CoV-2 S proteins bind with similar affinities to human ACE2 (19). In contrast, a modelling study suggested that binding of SARS-CoV-2 is stronger (20), which was convincingly confirmed by structural and biochemical data (16,17).…”

Section: Introductionmentioning

confidence: 83%

“…It cleaves both the SARS-CoV S protein and the virus receptor, ACE2, promoting both the viral uptake and the viral and cellular membrane fusion events (12)(13)(14). The critical residues contributing to the receptor-spike protein interaction were first determined for SARS-CoV-1 (15) and recently in three independent studies for SARS-CoV-2 (16)(17)(18). It has been proposed by biolayer interferometry studies that the receptorbinding domains of SARS-CoV-1 and SARS-CoV-2 S proteins bind with similar affinities to human ACE2 (19).…”

Section: Introductionmentioning

confidence: 99%

Bioinformatic characterization of angiotensin-converting enzyme 2, the entry receptor for SARS-CoV-2

Barker

Parkkila

2020

Preprint

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The World Health Organization declared the COVID-19 epidemic a public health emergency of international concern on March 11th, 2020, and the pandemic is rapidly spreading worldwide.COVID-19 is caused by a novel coronavirus SARS-CoV-2, which enters human target cells via angiotensin converting enzyme 2 (ACE2). We used a number of bioinformatics tools to computationally characterize ACE2 by determining its cell-specific expression, putative functions, and transcriptional regulation. The small intestine expressed higher levels of ACE2 than any other organ. The large intestine, kidney and testis showed moderate signals, whereas the signal was weak in the lung specimens. Single cell RNA-Seq data indicated positive signals along the respiratory tract in the key protective cell types including the goblet and ciliary epithelial cells, as well as in the endothelial cells and type I pneumocytes. Gene ontology analysis suggested that, besides its classical role in renin-angiotensin system, ACE2 may be functionally associated with angiogenesis/blood vessel morphogenesis. A novel tool for the prediction of transcription factor binding sites identified several putative sites for determined transcription factors within the ACE2 gene promoter. Our results also confirmed that age and gender play no significant role in the regulation of ACE2 mRNA expression in the lung.

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“…Once infection started, one monomer turned "up" its RBD to expose enough space to ACE2, inducing further conformational open and loose for proteolysis [1,3] . Atomic-level structural analysis suggested that the spatial interaction and interface between SARS-CoV-2 RBD and ACE2 was mostly in accordance with the SARS-CoV case [4] . Besides, a Cryo-EM structure of SARS-CoV-2 S protein trimer published recently showed that one of the three RBDs was in "up" conformation and naturally exposed the whole interaction interface [5] , while the classic closed symmetric trimer still existed [6] .…”

Section: Introductionmentioning

confidence: 83%

“…However, the real scenario is much more problematic. Several independent peer-reviewed studies as well as preprinted ones have proved that all structurally known SARS-CoV specific antibodies, including S230, 80R, m396 and F26G19, have no cross-reactivity of SARS-CoV-2 [4,5,9] . These antibodies all compete with ACE2 to bind SARS-CoV RBD, but their epitopes only have limited overlaps of the full ACE2-RBD interface, which could be the reason of lacking cross-reactivity.…”

Section: Discussionmentioning

confidence: 99%

Blocking antibodies against SARS-CoV-2 RBD isolated from a phage display antibody library using a competitive biopanning strategy

Zeng

Li²,

Lin³

et al. 2020

Preprint

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The infection of the novel coronavirus SARS-CoV-2 have caused more than 150,000 deaths, but no vaccine or specific therapeutic antibody is currently available. SARS-CoV-2 relies on its spike protein, in particular the receptor binding domain (RBD), to bind human cell receptor angiotensinconverting enzyme 2 (ACE2) for viral entry, and thus targeting RBD holds the promise for preventing SARS-CoV-2 infection. In this work, a competitive biopanning strategy of a phage display antibody library was applied to screen blocking antibodies against RBD. High-affinity antibodies were enriched after the first round using a standard panning process in which RBD-His recombinant protein was immobilized as a bait. At the next two rounds, immobilized ACE2-Fc and free RBD-His proteins were mixed with the enriched phage antibodies. Antibodies binding to RBD at epitopes different from ACE2-binding site were captured by the immobilized ACE2-Fc, forming a "sandwich" complex. Only antibodies competed with ACE2 for recognizing RBD at the same or similar epitopes can bind to the free RBD-His in the supernatant and be subsequently separated by the Ni-NTA magnetic beads. Top 1 lead from the competitive biopanning of a synthetic antibody library, Lib AB1, was produced as the full-length IgG1 format. It was proved to competitively block the binding of RBD to ACE2 protein, and potently inhibit SARS-CoV-2 pseudovirus infection of ACE2-overexpressing Hela cells with IC50 values of 12nM. Nevertheless, top 1 lead from the standard biopanning of Lib AB1, can only bind to RBD in vitro but not have the blocking or neutralization activity. Our strategy can efficiently isolate the blocking antibodies of RBD, and it would speed up the discovery of neutralizing antibodies against SARS-CoV-2.

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Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2

Abstract: Highlights d SARS-CoV-2 interacts with hACE2 via S protein CTD d A 2.5-Å structure of SARS-CoV-2-CTD in complex with hACE2 is resolved d The SARS-CoV-2-CTD displays stronger affinity for hACE2 compared with SARS-RBD d SARS-CoV-2 -CTD is antigenically different from SARS-RBD

Cited by 2,801 publications

References 57 publications

Cross-reactive neutralization of SARS-CoV-2 by serum antibodies from recovered SARS patients and immunized animals

Cross-reactive neutralization of SARS-CoV-2 by serum antibodies from recovered SARS patients and immunized animals

Bioinformatic characterization of angiotensin-converting enzyme 2, the entry receptor for SARS-CoV-2

Blocking antibodies against SARS-CoV-2 RBD isolated from a phage display antibody library using a competitive biopanning strategy

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