Host cell entry of Middle East respiratory syndrome coronavirus after two-step, furin-mediated activation of the spike protein

SARS-CoV-2 uses RBD of Spike (S) protein to attach with human cell via ACE2 receptor, followed by protease priming at S1/S2 site resulted in host cell entry and pathogenesis. In this context, we focused our aim in studying natural mutations harboring in Spike protein of SARS-CoV-2. We have analyzed 420 COVID-19 cases. G476S and V483G mutation are observed which lies in the RBD region where as the prevalent D614G mutation is observed in the vicinity of S1/S2 site. Interestingly MD simulation supports strong favorable interaction of ACE2 with RBD region containing V483A mutation as compared to G476S and reference wild Wuhan S protein. Radius of gyration analysis also showed high degree of compactness in V483A. The landscape plot and Gibbs free energy also support our findings. Overall, our study indicates that V483G in the RBD region can enhance its binding with the human ACE2 receptor. Interestingly D614G mutation in vicinity of S1/S2 region introduced a new cleavage site specific for a serine protease elastase that is anticipated to broaden the virus host cell tropism. Hence, both V483A and D614G mutations led to enhanced and broaden the virus host cell entry and transmission of the disease. Further epitope mapping analysis revealed G476S and D614G mutations as antigenic determinants and thus these mutations are important while designing a therapeutics vaccine or chimeric antibody. This finding will help in further understanding the role of such arising mutations in modulating immunogenicity, viral tropism and pathogenesis of the disease, which in lieu will help in designing vaccine more precisely to mitigate pandemic COVID-19.

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Mutations in Spike Protein of SARS-CoV-2 Modulate Receptor Binding, Membrane Fusion and Immunogenicity: An Insight into Viral Tropism and Pathogenesis of COVID-19

10.26434/chemrxiv.12320567 ◽

2020 ◽

Cited By ~ 1

Author(s):

Dr. Priyanka Saha ◽

Ranabir Majumder ◽

sourabrata chakraborty ◽

Amit Kumar Srivastava ◽

Mahitosh Mandal ◽

...

Keyword(s):

Host Cell ◽

Cell Entry ◽

Spike Protein ◽

Antigenic Determinants ◽

Chimeric Antibody ◽

Radius Of Gyration ◽

S Protein ◽

Viral Tropism ◽

Host Cell Entry ◽

High Degree

SARS-CoV-2 uses RBD of Spike (S) protein to attach with human cell via ACE2 receptor, followed by protease priming at S1/S2 site resulted in host cell entry and pathogenesis. In this context, we focused our aim in studying natural mutations harboring in Spike protein of SARS-CoV-2. We have analyzed 420 COVID-19 cases. G476S and V483G mutation are observed which lies in the RBD region where as the prevalent D614G mutation is observed in the vicinity of S1/S2 site. Interestingly MD simulation supports strong favorable interaction of ACE2 with RBD region containing V483A mutation as compared to G476S and reference wild Wuhan S protein. Radius of gyration analysis also showed high degree of compactness in V483A. The landscape plot and Gibbs free energy also support our findings. Overall, our study indicates that V483G in the RBD region can enhance its binding with the human ACE2 receptor. Interestingly D614G mutation in vicinity of S1/S2 region introduced a new cleavage site specific for a serine protease elastase that is anticipated to broaden the virus host cell tropism. Hence, both V483A and D614G mutations led to enhanced and broaden the virus host cell entry and transmission of the disease. Further epitope mapping analysis revealed G476S and D614G mutations as antigenic determinants and thus these mutations are important while designing a therapeutics vaccine or chimeric antibody. This finding will help in further understanding the role of such arising mutations in modulating immunogenicity, viral tropism and pathogenesis of the disease, which in lieu will help in designing vaccine more precisely to mitigate pandemic COVID-19.

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Faculty Opinions recommendation of Host cell entry by apicomplexa parasites requires actin polymerization in the host cell.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1157765.618988 ◽

2009 ◽

Author(s):

Marc Lecuit ◽

Serge Mostowy

Keyword(s):

Host Cell ◽

Actin Polymerization ◽

Cell Entry ◽

Host Cell Entry

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Faculty Opinions recommendation of Plasticity and redundancy among AMA-RON pairs ensure host cell entry of Toxoplasma parasites.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718453306.793496526 ◽

2014 ◽

Author(s):

Vernon Carruthers

Keyword(s):

Host Cell ◽

Cell Entry ◽

Host Cell Entry

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Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells

Journal of Virology ◽

10.1128/jvi.00683-18 ◽

2018 ◽

Vol 92 (19) ◽

Cited By ~ 30

Author(s):

Shutoku Matsuyama ◽

Kazuya Shirato ◽

Miyuki Kawase ◽

Yutaka Terada ◽

Kengo Kawachi ◽

...

Keyword(s):

Middle East ◽

Viral Entry ◽

Cathepsin L ◽

Inhibitory Effect ◽

Middle East Respiratory Syndrome ◽

Cell Entry ◽

Target Cells ◽

Furin Cleavage ◽

S Protein ◽

Entry Assay

ABSTRACT Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes host cellular proteases to enter cells. A previous report shows that furin, which is distributed mainly in the Golgi apparatus and cycled to the cell surface and endosomes, proteolytically activates the MERS-CoV spike (S) protein following receptor binding to mediate fusion between the viral and cellular membranes. In this study, we reexamined furin usage by MERS-CoV using a real-time PCR-based virus cell entry assay after inhibition of cellular proteases. We found that the furin inhibitor dec-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, dec-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2. Furthermore, a virus cell entry assay and a cell-cell fusion assay provided no evidence that the S protein was activated by exogenous furin. Therefore, we conclude that furin does not play a role in entry of MERS-CoV into cells and that the inhibitory effect of dec-RVKR-CMK is specific for TMPRSS2 and cathepsin L rather than furin. IMPORTANCE Previous studies using the furin inhibitor dec-RVKR-CMK suggest that MERS-CoV utilizes a cellular protease, furin, to activate viral glycoproteins during cell entry. However, we found that dec-RVKR-CMK inhibits not only furin but also other proteases. Furthermore, we found no evidence that MERS-CoV uses furin. These findings suggest that previous studies in the virology field based on dec-RVKR-CMK should be reexamined carefully. Here we describe appropriate experiments that can be used to assess the effect of protease inhibitors on virus cell entry.

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Middle East respiratory syndrome coronavirus (MERS-CoV) entry inhibitors targeting spike protein

Virus Research ◽

10.1016/j.virusres.2014.10.007 ◽

2014 ◽

Vol 194 ◽

pp. 200-210 ◽

Cited By ~ 49

Author(s):

Shuai Xia ◽

Qi Liu ◽

Qian Wang ◽

Zhiwu Sun ◽

Shan Su ◽

...

Keyword(s):

Middle East ◽

Middle East Respiratory Syndrome ◽

Spike Protein ◽

Entry Inhibitors

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Mapping the pH sensors critical for host cell entry by a complex Nonenveloped Virus

Access Microbiology ◽

10.1099/acmi.ac2019.po0259 ◽

2019 ◽

Vol 1 (1A) ◽

Author(s):

Weining Wu ◽

Polly Roy

Keyword(s):

Host Cell ◽

Cell Entry ◽

Ph Sensors ◽

Nonenveloped Virus ◽

Host Cell Entry

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A structure-based rationale for sialic acid independent host-cell entry of Sosuga virus

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1906717116 ◽

2019 ◽

Vol 116 (43) ◽

pp. 21514-21520 ◽

Cited By ~ 3

Author(s):

Alice J. Stelfox ◽

Thomas A. Bowden

Keyword(s):

Sialic Acid ◽

Host Cell ◽

Receptor Binding ◽

Cell Attachment ◽

Cell Entry ◽

Head Region ◽

Close Proximity ◽

Host Cell Attachment ◽

Host Cell Entry

The bat-borne paramyxovirus, Sosuga virus (SosV), is one of many paramyxoviruses recently identified and classified within the newly established genus Pararubulavirus, family Paramyxoviridae. The envelope surface of SosV presents a receptor-binding protein (RBP), SosV-RBP, which facilitates host-cell attachment and entry. Unlike closely related hemagglutinin neuraminidase RBPs from other genera of the Paramyxoviridae, SosV-RBP and other pararubulavirus RBPs lack many of the stringently conserved residues required for sialic acid recognition and hydrolysis. We determined the crystal structure of the globular head region of SosV-RBP, revealing that while the glycoprotein presents a classical paramyxoviral six-bladed β-propeller fold and structurally classifies in close proximity to paramyxoviral RBPs with hemagglutinin-neuraminidase (HN) functionality, it presents a receptor-binding face incongruent with sialic acid recognition. Hemadsorption and neuraminidase activity analysis confirms the limited capacity of SosV-RBP to interact with sialic acid in vitro and indicates that SosV-RBP undergoes a nonclassical route of host-cell entry. The close overall structural conservation of SosV-RBP with other classical HN RBPs supports a model by which pararubulaviruses only recently diverged from sialic acid binding functionality.

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Peptide-Protein Interaction Studies of Antimicrobial Peptides Targeting Middle East Respiratory Syndrome Coronavirus Spike Protein: An In Silico Approach

Advances in Bioinformatics ◽

10.1155/2019/6815105 ◽

2019 ◽

Vol 2019 ◽

pp. 1-16 ◽

Cited By ~ 5

Author(s):

Sabeena Mustafa ◽

Hanan Balkhy ◽

Musa Gabere

Keyword(s):

Middle East ◽

Antimicrobial Peptides ◽

Protein Interactions ◽

In Silico ◽

Therapeutic Option ◽

Binding Mode ◽

Middle East Respiratory Syndrome ◽

Spike Protein ◽

In Silico Docking ◽

Peptide Database

There is no effective therapeutic or vaccine for Middle East Respiratory Syndrome and this study attempts to find therapy using peptide by establishing a basis for the peptide-protein interactions through in silico docking studies for the spike protein of MERS-CoV. The antimicrobial peptides (AMPs) were retrieved from the antimicrobial peptide database (APD3) and shortlisted based on certain important physicochemical properties. The binding mode of the shortlisted peptides was measured based on the number of clusters which forms in a protein-peptide docking using Piper. As a result, we identified a list of putative AMPs which binds to the spike protein of MERS-CoV, which may be crucial in providing the inhibitory action. It is observed that seven putative peptides have good binding score based on cluster size cutoff of 208. We conclude that seven peptides, namely, AP00225, AP00180, AP00549, AP00744, AP00729, AP00764, and AP00223, could possibly have binding with the active site of the MERS-CoV spike protein. These seven AMPs could serve as a therapeutic option for MERS and enhance its treatment outcome.

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