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A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity.
Biochem Biophys Res Commun. 2006 May 26; 344(1):106-13.BB

Abstract

The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and cell-associated angiotensin-converting enzyme 2 (ACE2), the functional receptor for SARS-CoV and failed to block S protein-mediated pseudovirus entry, indicating that this point mutation also disrupted the receptor-binding motif (RBM) in the RBD. Taken together, these data provide direct evidence to show that a single amino acid residue at key position in the RBD can determine the major function of SARS-CoV S protein and imply for designing SARS vaccines and therapeutics.

Authors+Show Affiliations

Lindsley F. Kimball Research Institute, The New York Blood Center, New York, NY 10021, USA. yhe@nybloodcenter.orgNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16615996

Citation

He, Yuxian, et al. "A Single Amino Acid Substitution (R441A) in the Receptor-binding Domain of SARS Coronavirus Spike Protein Disrupts the Antigenic Structure and Binding Activity." Biochemical and Biophysical Research Communications, vol. 344, no. 1, 2006, pp. 106-13.
He Y, Li J, Jiang S. A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity. Biochem Biophys Res Commun. 2006;344(1):106-13.
He, Y., Li, J., & Jiang, S. (2006). A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity. Biochemical and Biophysical Research Communications, 344(1), 106-13.
He Y, Li J, Jiang S. A Single Amino Acid Substitution (R441A) in the Receptor-binding Domain of SARS Coronavirus Spike Protein Disrupts the Antigenic Structure and Binding Activity. Biochem Biophys Res Commun. 2006 May 26;344(1):106-13. PubMed PMID: 16615996.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity. AU - He,Yuxian, AU - Li,Jingjing, AU - Jiang,Shibo, Y1 - 2006/03/30/ PY - 2006/03/15/received PY - 2006/03/21/accepted PY - 2006/4/18/pubmed PY - 2006/6/24/medline PY - 2006/4/18/entrez SP - 106 EP - 13 JF - Biochemical and biophysical research communications JO - Biochem Biophys Res Commun VL - 344 IS - 1 N2 - The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and cell-associated angiotensin-converting enzyme 2 (ACE2), the functional receptor for SARS-CoV and failed to block S protein-mediated pseudovirus entry, indicating that this point mutation also disrupted the receptor-binding motif (RBM) in the RBD. Taken together, these data provide direct evidence to show that a single amino acid residue at key position in the RBD can determine the major function of SARS-CoV S protein and imply for designing SARS vaccines and therapeutics. SN - 0006-291X UR - https://www.unboundmedicine.com/medline/citation/16615996/A_single_amino_acid_substitution__R441A__in_the_receptor_binding_domain_of_SARS_coronavirus_spike_protein_disrupts_the_antigenic_structure_and_binding_activity_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(06)00678-4 DB - PRIME DP - Unbound Medicine ER -