Abstract
The coronavirus spike (S) protein mediates infection of receptor-expressing host cells and is a critical target for antiviral neutralizing antibodies. Angiotensin-converting enzyme 2 (ACE2) is a functional receptor for the coronavirus (severe acute respiratory syndrome (SARS)-CoV) that causes SARS. Here we demonstrate that a 193-amino acid fragment of the S protein (residues 318-510) bound ACE2 more efficiently than did the full S1 domain (residues 12-672). Smaller S protein fragments, expressing residues 327-510 or 318-490, did not detectably bind ACE2. A point mutation at aspartic acid 454 abolished association of the full S1 domain and of the 193-residue fragment with ACE2. The 193-residue fragment blocked S protein-mediated infection with an IC(50) of less than 10 nm, whereas the IC(50) of the S1 domain was approximately 50 nm. These data identify an independently folded receptor-binding domain of the SARS-CoV S protein.
MeSH Terms
Alanine/chemistry
Angiotensin-Converting Enzyme 2
Carboxypeptidases/chemistry,metabolism
Cell Line
Cysteine/chemistry
Dose-Response Relationship, Drug
Electrophoresis, Polyacrylamide Gel
Flow Cytometry
Glutamic Acid/chemistry
Humans
Inhibitory Concentration 50
Membrane Glycoproteins/chemistry
Mutation
Peptidyl-Dipeptidase A
Point Mutation
Protein Binding
Protein Structure, Tertiary
SARS Virus/metabolism
Spike Glycoprotein, Coronavirus
Transfection
Viral Envelope Proteins/chemistry
Chemicals
Membrane Glycoproteins
Spike Glycoprotein, Coronavirus
Viral Envelope Proteins
spike glycoprotein, SARS-CoV
spike protein, mouse hepatitis virus
Glutamic Acid
Carboxypeptidases
Peptidyl-Dipeptidase A
ACE2 protein, human
Angiotensin-Converting Enzyme 2
Cysteine
Alanine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wong Swee Kee
Partners AIDS Research Center, Brigham and Women's Hospital, Department of Medicine (Microbiology and Molecular Genetics), Harvard Medical School, Boston, Massachusetts 02115, USA.
Li Wenhui
Moore Michael J
Choe Hyeryun
Farzan Michael
References (18)
18 references, click to expand
-
Newly discovered coronavirus as the primary cause of severe acute respiratory syndrome.
Lancet. 2003 Jul 26;362(9380):263-70
PMID: 12892955
-
Coronavirus spike proteins in viral entry and pathogenesis.
Virology. 2001 Jan 20;279(2):371-4
PMID: 11162792
-
Tyrosine sulfation of the amino terminus of CCR5 facilitates HIV-1 entry.
Cell. 1999 Mar 5;96(5):667-76
PMID: 10089882
-
Identification of a novel coronavirus in patients with severe acute respiratory syndrome.
N Engl J Med. 2003 May 15;348(20):1967-76
PMID: 12690091
-
Localization of neutralizing epitopes and the receptor-binding site within the amino-terminal 330 amino acids of the murine coronavirus spike protein.
J Virol. 1994 Sep;68(9):5403-10
PMID: 7520090
-
Cloning of the mouse hepatitis virus (MHV) receptor: expression in human and hamster cell lines confers susceptibility to MHV.
J Virol. 1991 Dec;65(12):6881-91
PMID: 1719235
-
Human coronavirus 229E: receptor binding domain and neutralization by soluble receptor at 37 degrees C.
J Virol. 2003 Apr;77(7):4435-8
PMID: 12634402
-
Several members of the mouse carcinoembryonic antigen-related glycoprotein family are functional receptors for the coronavirus mouse hepatitis virus-A59.
J Virol. 1993 Jan;67(1):1-8
PMID: 8380065
-
The Genome sequence of the SARS-associated coronavirus.
Science. 2003 May 30;300(5624):1399-404
PMID: 12730501
-
Human aminopeptidase N is a receptor for human coronavirus 229E.
Nature. 1992 Jun 4;357(6377):420-2
PMID: 1350662
-
Identification of a receptor-binding domain of the spike glycoprotein of human coronavirus HCoV-229E.
J Virol. 2003 Feb;77(4):2530-8
PMID: 12551991
-
A novel coronavirus associated with severe acute respiratory syndrome.
N Engl J Med. 2003 May 15;348(20):1953-66
PMID: 12690092
-
Proteolytic cleavage of peplomeric glycoprotein E2 of MHV yields two 90K subunits and activates cell fusion.
Adv Exp Med Biol. 1984;173:25-35
PMID: 6331116
-
Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus.
Nature. 2003 Nov 27;426(6965):450-4
PMID: 14647384
-
Aetiology: Koch's postulates fulfilled for SARS virus.
Nature. 2003 May 15;423(6937):240
PMID: 12748632
-
The level of CD4 expression limits infection of primary rhesus monkey macrophages by a T-tropic simian immunodeficiency virus and macrophagetropic human immunodeficiency viruses.
J Virol. 2000 Dec;74(23):10984-93
PMID: 11069993
-
Characterization of a novel coronavirus associated with severe acute respiratory syndrome.
Science. 2003 May 30;300(5624):1394-9
PMID: 12730500
-
Spike glycoprotein cleavage recognition site analysis of infectious bronchitis virus.
Avian Dis. 2001 Apr-Jun;45(2):366-72
PMID: 11417816