Molecular modelling of S1 and S2 subunits of SARS coronavirus spike glycoprotein

Spiga, Ottavia; Bernini, Andrea; Ciutti, Arianna; Chiellini, S.; Menciassi, N.; Finetti, Francesca; Causarono, Vincenza; Anselmi, Francesca; Prischi, Filippo; Niccolai, Neri

doi:10.1016/j.bbrc.2003.08.122

Cited by 87 publications

(94 citation statements)

References 20 publications

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“…Two functional domains at the amino (S1) and carboxy (S2) termini of the S protein are conserved among the coronaviruses. The S1 and S2 domain of SARS-CoV S protein can be identified by sequence alignment with other coronavirus S proteins, especially with the more conserved S2 domain (8)(9)(10). The S protein is also the major antigenic determinant for coronaviruses (9,(11)(12)(13)(14).…”

mentioning

confidence: 99%

Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association

Sui

Murakami

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

567

603

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Effective prophylaxis and antiviral therapies are urgently needed in the event of reemergence of the highly contagious and often fatal severe acute respiratory syndrome (SARS) coronavirus (SARSCoV) infection. We have identified eight recombinant human single-chain variable region fragments (scFvs) against the S1 domain of spike (S) protein of the SARS-CoV from two nonimmune human antibody libraries. One scFv 80R efficiently neutralized SARS-CoV and inhibited syncytia formation between cells expressing the S protein and those expressing the SARS-CoV receptor angiotensin-converting enzyme 2 (ACE2). Mapping of the 80R epitope showed it is located within the N-terminal 261-672 amino acids of S protein and is not glycosylation-dependent. 80R scFv competed with soluble ACE2 for association with the S1 domain and bound S1 with high affinity (equilibrium dissociation constant, K d ‫؍‬ 32.3 nM). A human IgG1 form of 80R bound S1 with a 20-fold higher affinity of 1.59 nM comparable to that of ACE2 (K d ‫؍‬ 1.70 nM), and neutralized virus 20-fold more efficiently than the 80R scFv. These data suggest that the 80R human monoclonal antibody may be a useful viral entry inhibitor for the emergency prophylaxis and treatment of SARS, and that the ACE2-binding site of S1 could be an attractive target for subunit vaccine and drug development.T he severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV), a newly emergent member in the family Coronaviridae, causes SARS for which there are no vaccines or effective therapies currently available (1-4). It has been reported that high titers of protecting IgG antibody to SARS-CoV are present in convalescent serum, and SARS patients show clinical improvement if they are given serum from previously infected patients (5, 6). These observations suggest that passive immunization with human monoclonal antibodies could be developed for the treatment of SARS (7). The spike (S) proteins of coronaviruses are large type-I transmembrane glycoproteins that are responsible for receptor binding and membrane fusion. Two functional domains at the amino (S1) and carboxy (S2) termini of the S protein are conserved among the coronaviruses. The S1 and S2 domain of SARS-CoV S protein can be identified by sequence alignment with other coronavirus S proteins, especially with the more conserved S2 domain (8-10). The S protein is also the major antigenic determinant for coronaviruses (9,(11)(12)(13)(14). It has recently been demonstrated that the binding of the S1 domain to its receptor angiotensinconverting enzyme 2 (ACE2) on host cells is responsible for SARS-CoV entry into cells (15). Therefore, we targeted the S1 protein for generation of neutralizing human monoclonal antibodies. Here we report the identification, production, and characterization of a neutralizing human monoclonal antibody 80R against SARS-CoV that blocks the binding of S1 to ACE2. Materials and MethodsExpression and Purification of SARS-CoV S1 and Truncated S1. Plasmids encoding SARS-CoV S protein residues 12-672, 12-327, o...

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mentioning

confidence: 99%

Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association

Sui

Murakami

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

567

603

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“…Molecular modeling has proposed a hypothetic division of these two subunits between Leu681-Asp727 ( Fig. 1A) (18). S1 was further divided into S1.1 (Ser12-Thr535) and S1.2 (Gly534-Ser798).…”

mentioning

confidence: 99%

Identification of Two Neutralizing Regions on the Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein Produced from the Mammalian Expression System

Wang

Chou

Sakhatskyy

et al. 2005

J Virol

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The Spike (S) protein of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) plays important roles in viral pathogenesis and potentially in the development of an effective vaccine against this virulent infectious disease. In this study, the codon-optimized S gene of SARS-CoV was synthesized to construct DNA vaccine plasmids expressing either the full-length or segments of the S protein. High titer S-specific immunoglobulin G antibody responses were elicited in rabbits immunized with DNA against various segments of the S protein. Two neutralizing domains were identified on the S protein, one at the N terminus (Ser12-Thr535) and the other near the C terminus (Arg797-Ile1192).

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“…Ectodomain S1 presents a region of hypervariability (4). Exchange of aminoacids in such an important region for coronavirus antigenicity is greatly used in the study of SARS, because most mutations occur in this subunity (17,22).…”

Section: Discussionmentioning

confidence: 99%

Isolation of bovine coronavirus (BCoV) in monolayers of HmLu-1 cells

Jerez¹,

Gregori²,

Brandão³

et al. 2005

Braz. J. Microbiol.

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Isolation of BCoV was performed in monolayers of HmLu-1 cells, using 20 fecal samples from clinical cases of diarrhea in calves. Samples were positive for BCoV by means of hemagglutination / hemagglutination inhibition (HA/HI). Up to the fifth serial passage, 13 of these isolates presented syncytial cytopathic effect, similar to Kakegawa standard strain. When isolates were submitted to seroneutralization with gammaglobulin antibovine coronavirus, 8 of them were considered to be positive, once cytopathic effect was neutralized. After titration and seroneutralization in microplates, only three of them were confirmed as positive. The lineage HmLu-1 showed higher permissivity to BCoV isolation, but the low intensity of viral replication demonstrated that new methodologies should be developed for this purpose and then submitted to confirmation of isolation by means of seroneutralization.

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Molecular modelling of S1 and S2 subunits of SARS coronavirus spike glycoprotein

Cited by 87 publications

References 20 publications

Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association

Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association

Identification of Two Neutralizing Regions on the Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein Produced from the Mammalian Expression System

Isolation of bovine coronavirus (BCoV) in monolayers of HmLu-1 cells

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