The use of hepatitis C virus NS3/4A and secreted alkaline phosphatase to quantitate cell–cell membrane fusion mediated by severe acute respiratory syndrome coronavirus S protein and the receptor angiotensin-converting enzyme 2

SUMMARYThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein mediates infection of cells expressing angiotensin-converting enzyme 2 (ACE2). ACE2 is also the viral receptor of SARS-CoV (SARS-CoV-1), a related coronavirus that emerged in 2002-2003. Horseshoe bats (genus Rhinolophus) are presumed to be the original reservoir of both viruses, and a SARS-like coronavirus, RaTG13, closely related SARS-CoV-2, has been isolated from one horseshoe-bat species. Here we characterize the ability of S-protein receptor-binding domains (RBDs) of SARS-CoV-1, SARS-CoV-2, and RaTG13 to bind a range of ACE2 orthologs. We observed that the SARS-CoV-2 RBD bound human, pangolin, and horseshoe bat (R. macrotis) ACE2 more efficiently than the SARS-CoV-1 or RaTG13 RBD. Only the RaTG13 RBD bound rodent ACE2 orthologs efficiently. Five mutations drawn from ACE2 orthologs of nine Rhinolophus species enhanced human ACE2 binding to the SARS-CoV-2 RBD and neutralization of SARS-CoV-2 by an immunoadhesin form of human ACE2 (ACE2-Fc). Two of these mutations impaired neutralization of SARS-CoV-1. An ACE2-Fc variant bearing all five mutations neutralized SARS-CoV-2 five-fold more efficiently than human ACE2-Fc. These data narrow the potential SARS-CoV-2 reservoir, suggest that SARS-CoV-1 and -2 originate from distinct bat species, and identify a more potently neutralizing form of ACE2-Fc.

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Cooperative multivalent receptor binding promotes exposure of the SARS-CoV-2 fusion machinery core

10.1101/2021.05.24.445443 ◽

2021 ◽

Author(s):

Alexander J Pak ◽

Alvin Yu ◽

Zunlong Ke ◽

John Briggs ◽

Gregory A Voth

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Membrane Fusion ◽

Receptor Binding ◽

Coarse Grained ◽

Viral Fusion ◽

Converting Enzyme ◽

Angiotensin Converting Enzyme 2 ◽

Viral Binding ◽

Molecular Events

The molecular events that permit the spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to bind, fuse, and enter cells are important to understand for both fundamental and therapeutic reasons. Spike proteins consist of S1 and S2 domains, which recognize angiotensin-converting enzyme 2 (ACE2) receptors and contain the viral fusion machinery, respectively. Ostensibly, the binding of spike trimers to ACE2 receptors promotes the preparation of the fusion machinery by dissociation of the S1 domains. We report the development and use of coarse-grained models and simulations to investigate the dynamical mechanisms involved in viral binding and exposure of the S2 trimeric core. We show that spike trimers cooperatively bind to multiple ACE2 dimers. The multivalent interaction cyclically and processively induces S1 dissociation, thereby exposing the S2 core containing the fusion machinery. Our simulations thus reveal an important concerted interaction between spike trimers and ACE2 dimers that primes the virus for membrane fusion and entry.

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Naturally mutated spike proteins of SARS-CoV-2 variants show differential levels of cell entry

10.1101/2020.06.15.151779 ◽

2020 ◽

Cited By ~ 13

Author(s):

Seiya Ozono ◽

Yanzhao Zhang ◽

Hirotaka Ode ◽

Toong Seng Tan ◽

Kazuo Imai ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Causative Agent ◽

Cell Entry ◽

Viral Infectivity ◽

Converting Enzyme ◽

S Protein ◽

Angiotensin Converting Enzyme 2 ◽

Naturally Occurring ◽

Spike Proteins

AbstractThe causative agent of the coronavirus disease 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is steadily mutating during continuous transmission among humans. Such mutations can occur in the spike (S) protein that binds to the angiotensin-converting enzyme-2 (ACE2) receptor and is cleaved by transmembrane protease serine 2 (TMPRSS2). However, whether S mutations affect SARS-CoV-2 infectivity remains unknown. Here, we show that naturally occurring S mutations can reduce or enhance cell entry via ACE2 and TMPRSS2. A SARS-CoV-2 S-pseudotyped lentivirus exhibits substantially lower entry than SARS-CoV S. Among S variants, the D614G mutant shows the highest cell entry, as supported by structural observations. Nevertheless, the D614G mutant remains susceptible to neutralization by antisera against prototypic viruses. Taken together, these data indicate that the D614G mutation enhances viral infectivity while maintaining neutralization susceptibility.

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Interaction of severe acute respiratory syndrome-coronavirus and NL63 coronavirus spike proteins with angiotensin converting enzyme-2

Journal of General Virology ◽

10.1099/vir.0.2008/003962-0 ◽

2008 ◽

Vol 89 (11) ◽

pp. 2741-2745 ◽

Cited By ~ 31

Author(s):

Alison C. Mathewson ◽

Alexandra Bishop ◽

Yongxiu Yao ◽

Fred Kemp ◽

Junyuan Ren ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Converting Enzyme ◽

Severe Respiratory Distress ◽

S Protein ◽

Angiotensin Converting Enzyme 2 ◽

Lower Affinity ◽

Common Receptor ◽

Affinity Interaction ◽

Receptor Binding Protein

Although in different groups, the coronaviruses severe acute respiratory syndrome-coronavirus (SARS-CoV) and NL63 use the same receptor, angiotensin converting enzyme (ACE)-2, for entry into the host cell. Despite this common receptor, the consequence of entry is very different; severe respiratory distress in the case of SARS-CoV but frequently only a mild respiratory infection for NL63. Using a wholly recombinant system, we have investigated the ability of each virus receptor-binding protein, spike or S protein, to bind to ACE-2 in solution and on the cell surface. In both assays, we find that the NL63 S protein has a weaker interaction with ACE-2 than the SARS-CoV S protein, particularly in solution binding, but the residues required for contact are similar. We also confirm that the ACE-2-binding site of NL63 S lies between residues 190 and 739. A lower-affinity interaction with ACE-2 might partly explain the different pathological consequences of infection by SARS-CoV and NL63.

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Amino Acid Substitutions in the S2 Region Enhance Severe Acute Respiratory Syndrome Coronavirus Infectivity in Rat Angiotensin-Converting Enzyme 2-Expressing Cells

Journal of Virology ◽

10.1128/jvi.01143-07 ◽

2007 ◽

Vol 81 (19) ◽

pp. 10831-10834 ◽

Cited By ~ 7

Author(s):

Shuetsu Fukushi ◽

Tetsuya Mizutani ◽

Kouji Sakai ◽

Masayuki Saijo ◽

Fumihiro Taguchi ◽

...

Keyword(s):

Amino Acid ◽

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Host Species ◽

Molecular Basis ◽

Amino Acid Substitutions ◽

Converting Enzyme ◽

S Protein ◽

Angiotensin Converting Enzyme 2 ◽

Virus Adaptation

ABSTRACT To clarify the molecular basis of severe acute respiratory syndrome coronavirus (SARS-CoV) adaptation to different host species, we serially passaged SARS-CoV in rat angiotensin-converting enzyme 2 (ACE2)-expressing cells. After 15 passages, the virus (Rat-P15) came to replicate effectively in rat ACE2-expressing cells. Two amino acid substitutions in the S2 region were found on the Rat-P15 S gene. Analyses of the infectivity of the pseudotype-bearing S protein indicated that the two substitutions in the S2 region, especially the S950F substitution, were responsible for efficient infection. Therefore, virus adaptation to different host species can be induced by amino acid substitutions in the S2 region.

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Structural Similarity Analysis of the Spike Protein of SARS-CoV-2 and Other SARS-related Coronaviruses

10.20944/preprints202003.0409.v1 ◽

2020 ◽

Author(s):

YoungJoon Park ◽

Ju Won Ahn ◽

Sojung Hwang ◽

Kyoung Su Sung ◽

Jaejoon Lim ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Structural Similarity ◽

Spike Protein ◽

Similarity Analysis ◽

Converting Enzyme ◽

S Protein ◽

Angiotensin Converting Enzyme 2 ◽

Strong Affinity ◽

High Infectivity

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has high infectivity in humans, attributed to the strong affinity of its spike (S) protein to angiotensin-converting enzyme 2. Here, we analysed the structural similarity of the S protein between SARS-CoV-2 and other SARS-related coronaviruses. The S1 domain of the unclassified coronavirus RaTG13 was structurally very similar to that of SARS-CoV-2, implying that RaTG13 could be the origin of SARS-CoV-2.

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Retroviruses Pseudotyped with the Severe Acute Respiratory Syndrome Coronavirus Spike Protein Efficiently Infect Cells Expressing Angiotensin-Converting Enzyme 2

Journal of Virology ◽

10.1128/jvi.78.19.10628-10635.2004 ◽

2004 ◽

Vol 78 (19) ◽

pp. 10628-10635 ◽

Cited By ~ 130

Author(s):

Michael J. Moore ◽

Tatyana Dorfman ◽

Wenhui Li ◽

Swee Kee Wong ◽

Yanhan Li ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Fluorescent Protein ◽

Leukemia Virus ◽

Converting Enzyme ◽

Protein Variant ◽

Live Virus ◽

S Protein ◽

Angiotensin Converting Enzyme 2 ◽

Immunodeficiency Virus

ABSTRACT Infection of receptor-bearing cells by coronaviruses is mediated by their spike (S) proteins. The coronavirus (SARS-CoV) that causes severe acute respiratory syndrome (SARS) infects cells expressing the receptor angiotensin-converting enzyme 2 (ACE2). Here we show that codon optimization of the SARS-CoV S-protein gene substantially enhanced S-protein expression. We also found that two retroviruses, simian immunodeficiency virus (SIV) and murine leukemia virus, both expressing green fluorescent protein and pseudotyped with SARS-CoV S protein or S-protein variants, efficiently infected HEK293T cells stably expressing ACE2. Infection mediated by an S-protein variant whose cytoplasmic domain had been truncated and altered to include a fragment of the cytoplasmic tail of the human immunodeficiency virus type 1 envelope glycoprotein was, in both cases, substantially more efficient than that mediated by wild-type S protein. Using S-protein-pseudotyped SIV, we found that the enzymatic activity of ACE2 made no contribution to S-protein-mediated infection. Finally, we show that a soluble and catalytically inactive form of ACE2 potently blocked infection by S-protein-pseudotyped retrovirus and by SARS-CoV. These results permit studies of SARS-CoV entry inhibitors without the use of live virus and suggest a candidate therapy for SARS.

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Molecular Recognition of SARS-CoV-2 Spike Glycoprotein: Quantum Chemical Hot Spot and Epitope Analyses

Chemical Science ◽

10.1039/d0sc06528e ◽

2021 ◽

Author(s):

Chiduru Watanabe ◽

Yoshio Okiyama ◽

Shigenori Tanaka ◽

Kaori Fukuzawa ◽

Teruki Honma

Keyword(s):

Molecular Recognition ◽

Severe Acute Respiratory Syndrome ◽

Angiotensin Converting Enzyme ◽

Quantum Chemical ◽

Hot Spot ◽

Complex Structures ◽

Converting Enzyme ◽

Spike Glycoprotein ◽

S Protein ◽

Angiotensin Converting Enzyme 2

Due to the COVID-19 pandemic, researchers have attempted to identify complex structures of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike glycoprotein (S-protein) with angiotensin-converting enzyme 2 (ACE2) or...

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In-silico drug repurposing study: Amprenavir, enalaprilat, and plerixafor, potential drugs for destabilizing the SARS-CoV-2 S-protein-angiotensin-converting enzyme 2 complex

Results in Chemistry ◽

10.1016/j.rechem.2020.100094 ◽

2020 ◽

pp. 100094

Author(s):

Ivonne Buitrón-González ◽

Giovanny Aguilera-Durán ◽

Antonio Romo-Mancillas

Keyword(s):

Angiotensin Converting Enzyme ◽

In Silico ◽

Drug Repurposing ◽

Converting Enzyme ◽

S Protein ◽

Angiotensin Converting Enzyme 2

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Do sex-specific immunobiological factors and differences in angiotensin converting enzyme 2 (ACE2) expression explain increased severity and mortality of COVID-19 in males?

Diagnosis ◽

10.1515/dx-2020-0054 ◽

2020 ◽

Vol 7 (4) ◽

pp. 385-386 ◽

Cited By ~ 3

Author(s):

Jens Vikse ◽

Giuseppe Lippi ◽

Brandon Michael Henry

Keyword(s):

Mortality Rate ◽

Severe Acute Respiratory Syndrome ◽

Lung Injury ◽

Angiotensin Converting Enzyme ◽

Converting Enzyme ◽

Sars Coronavirus ◽

Immunomodulatory Treatment ◽

Angiotensin Converting Enzyme 2 ◽

Cytopathic Effects ◽

Severe Lung

AbstractCoronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome (SARS) coronavirus 2 (SARS-CoV-2), shares similarities with the former SARS outbreak, which was caused by SARS-CoV-1. SARS was characterized by severe lung injury due to virus-induced cytopathic effects and dysregulated hyperinflammatory state. COVID-19 has a higher mortality rate in men both inside and outside China. In this opinion paper, we describe how sex-specific immunobiological factors and differences in angiotensin converting enzyme 2 (ACE2) expression may explain the increased severity and mortality of COVID-19 in males. We highlight that immunomodulatory treatment must be tailored to the underlying immunobiology at different stages of disease. Moreover, by investigating sex-based immunobiological differences, we may enhance our understanding of COVID-19 pathophysiology and facilitate improved immunomodulatory strategies.

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