Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus

Li, Wenhui; Moore, Michael J.; Vasilieva, Natalya; Sui, Jianhua; Wong, Sharon; Berne, Michael A.; Somasundaran, Mohan; Sullivan, John L.; Luzuriaga, Katherine; Greenough, Thomas C.; Choe, Hyeryun; Farzan, Michael

doi:10.1038/nature02145

Cited by 5,718 publications

(5,701 citation statements)

References 31 publications

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“…We produced and purified a recombinant glycoprotein (similar to that described in Ref. 10) for biochemical studies because the soluble S1 antigen could not be isolated from a virus preparation. This form of recombinant protein was used for the original description of ACE2 as the virus receptor (10) and in a number of subsequent studies examining binding activity, folding, and even the role of the disulfides of S1 (32,33).…”

Section: Resultsmentioning

confidence: 99%

Significant Redox Insensitivity of the Functions of the SARS-CoV Spike Glycoprotein

Lavillette

Barbouche

Yao

et al. 2006

Journal of Biological Chemistry

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The capacity of the surface glycoproteins of enveloped viruses to mediate virus/cell binding and membrane fusion requires a proper thiol/disulfide balance. Chemical manipulation of their redox state using reducing agents or free sulfhydryl reagents affects virus/cell interaction. Conversely, natural thiol/disulfide rearrangements often occur during the cell interaction to trigger fusogenicity, hence the virus entry. We examined the relationship between the redox state of the 20 cysteine residues of the SARS-CoV (severe acute respiratory syndrome coronavirus) Spike glycoprotein S1 subdomain and its functional properties. Mature S1 exhibited ϳ4 unpaired cysteines, and chemically reduced S1 displaying up to ϳ6 additional unpaired cysteines still bound ACE2 and enabled fusion. In addition, virus/cell membrane fusion occurred in the presence of sulfhydryl-blocking reagents and oxidoreductase inhibitors. Thus, in contrast to various viruses including HIV (human immunodeficiency virus) examined in parallel, the functions of the SARS-CoV Spike glycoprotein exhibit a significant and surprising independence of redox state, which may contribute to the wide host range of the virus. These data suggest clues for molecularly engineering vaccine immunogens.SARS 3 -CoV is a new human coronavirus that causes a severe acute respiratory syndrome, sometimes with a fatal outcome. Its Spike glycoprotein is the major surface antigen and induces potent neutralizing antibodies (1-8). It is made up of S1 and S2 subdomains (2, 3). The S1 region interacts with angiotensin-converting enzyme 2 (ACE2), the primary cellular receptor (9 -12), L-SIGN (13), and L-SECtin (14), whereas S2 mediates membrane fusion (2, 3). Here, we investigated the functional role of the 20 conserved cysteine residues of mature S1.A variety of evidence has shown that the capacity of the viral envelope glycoproteins to mediate virus/cell membrane fusion depends on a precise thiol/disulfide balance within the viral surface complex (15-29). On one hand, manipulation of the native disulfide network of mature envelopes at the virus surface using reducing or alkylating reagents has important consequences for their subsequent capacity to carry out the virus/cell interaction process (15, 16, 19, 20, 23, 24, 26, and 28). On the other hand, natural and specific thiol/disulfide rearrangements occur within several viral envelopes to trigger conformational changes that insert the fusion peptide into the cell surface leading to virus entry (20,21,23,(25)(26)(27). For HIV, gentle chemical reduction of Env efficiently prevents viral infectivity by inhibiting the capacity of the surface subunit (SU) to bind its ligands on the target cell surface (15,19). In contrast, after receptor binding, fusion mediated by the transmembrane subunit occurs solely following reduction of SU by a cell-surface protein-disulfide isomerase (PDI) activity (19,(25)(26)(27). For these reasons, reducing or free sulfhydryl reagents and nonpermeant inhibitors of PDI (e.g. the thiol reagent DTNB and bacitracin, ...

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Section: Resultsmentioning

confidence: 99%

Significant Redox Insensitivity of the Functions of the SARS-CoV Spike Glycoprotein

Lavillette

Barbouche

Yao

et al. 2006

Journal of Biological Chemistry

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“…This protein contains two functional domains, S1 and S2, which are located in the N-and Cterminal regions, respectively. It has been previously demonstrated that the angiotensin-converting enzyme 2 on the cell surface functions as a receptor for the SARS-CoV S protein (27,28). A 193-aa small fragment within the S1 domain (S318 -510) was identified as a minimal receptor domain and contained multiple conformationdependent epitopes that induce highly potent neutralizing Abs (29,30).…”

mentioning

confidence: 99%

Screening and Identification of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific CTL Epitopes

Zhou

De-sheng

et al. 2006

The Journal of Immunology

107

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Severe acute respiratory syndrome (SARS) is a highly contagious and life-threatening disease that emerged in China in November 2002. A novel SARS-associated coronavirus was identified as its principal etiologic agent; however, the immunopathogenesis of SARS and the role of special CTLs in virus clearance are still largely uncharacterized. In this study, potential HLA-A*0201-restricted spike (S) and nucleocapsid protein-derived peptides were selected from an online database and screened for potential CTL epitopes by in vitro refolding and T2 cell-stabilization assays. The antigenicity of nine peptides which could refold with HLA-A*0201 molecules was assessed with an IFN-γ ELISPOT assay to determine the capacity to stimulate CTLs from PBMCs of HLA-A2+ SARS-recovered donors. A novel HLA-A*0201-restricted decameric epitope P15 (S411–420, KLPDDFMGCV) derived from the S protein was identified and found to localize within the angiotensin-converting enzyme 2 receptor-binding region of the S1 domain. P15 could significantly enhance the expression of HLA-A*0201 molecules on the T2 cell surface, stimulate IFN-γ-producing CTLs from the PBMCs of former SARS patients, and induce specific CTLs from P15-immunized HLA-A2.1 transgenic mice in vivo. Furthermore, significant P15-specific CTLs were induced from HLA-A2.1-transgenic mice immunized by a DNA vaccine encoding the S protein; suggesting that P15 was a naturally processed epitope. Thus, P15 may be a novel SARS-associated coronavirus-specific CTL epitope and a potential target for characterization of virus control mechanisms and evaluation of candidate SARS vaccines.

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“…The SARS-CoV S is quite divergent from those of other CoVs, exhibiting only 20-27% overall amino acid identity (5). Recent studies indicated that the SARS-CoV S is expressed as a noncleaved glycoprotein with an apparent mass of 180-200 kDa that interacts with a functional receptor identified as angiotensin-converting enzyme 2 (12,13).…”

mentioning

confidence: 99%

Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice

Bisht

Roberts

Vogel

et al. 2004

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

403

443

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Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus

Cited by 5,718 publications

References 31 publications

Significant Redox Insensitivity of the Functions of the SARS-CoV Spike Glycoprotein

Significant Redox Insensitivity of the Functions of the SARS-CoV Spike Glycoprotein

Screening and Identification of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific CTL Epitopes

Severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice

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