Localization of neutralizing epitopes and the receptor-binding site within the amino-terminal 330 amino acids of the murine coronavirus spike protein.

1994 ◽  
Vol 68 (9) ◽  
pp. 5403-5410 ◽  
Author(s):  
H Kubo ◽  
Y K Yamada ◽  
F Taguchi
Keyword(s):  
Amino Acids ◽  
Binding Site ◽  
Receptor Binding ◽  
Spike Protein ◽  
Receptor Binding Site ◽  
Amino Terminal ◽  
Neutralizing Epitopes ◽  
Murine Coronavirus

scholarly journals Evidence for the location of the receptor-binding site of human erythropoietin at the carboxyl-terminal domain

Blood ◽  
1991 ◽  
Vol 77 (6) ◽  
pp. 1203-1210 ◽  
Author(s):  
MR Fibi ◽  
W Stuber ◽  
P Hintz-Obertreis ◽  
G Luben ◽  
D Krumwieh ◽  
...  
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Terminal Region ◽  
Biologically Active ◽  
Receptor Binding Site ◽  
Specific Antibodies ◽  
Amino Terminal ◽  
Human Erythropoietin ◽  
A Cell ◽  
Carboxyl Terminal

Abstract Five different peptides (P1: 84′95; P2: 152′166; P3: 52′63; P4: 7′23; P5: 110′123) homologous to relatively hydrophilic regions of human erythropoietin (huEpo) have been synthesized to identify biologically active domains of the hormone. All peptides were able to induce high titers of peptide-specific antibodies in rabbits. Antisera from rabbits induced by recombinant huEpo (rhuEpo) contained a relatively high amount of antibodies preferentially directed against three peptides (P2, P4, and P5), of which P4 comprised the amino-terminal region, P2 the carboxyl-terminus, and P5 an interior region previously described as the receptor-binding site. The same three peptides were able to induce rhuEpo-specific antibodies, whereas P1 and P3 lacked this activity. Only peptide-P2-induced antisera inhibited the biologic activity of rhuEpo in a cell proliferation assay, indicating that the carboxyl-terminal region of the molecule is essentially involved in the biologic function of rhuEpo.

scholarly journals The Murine Coronavirus Hemagglutinin-esterase Receptor-binding Site: A Major Shift in Ligand Specificity through Modest Changes in Architecture

2012 ◽  
Vol 8 (1) ◽  
pp. e1002492 ◽  
Author(s):  
Martijn A. Langereis ◽  
Qinghong Zeng ◽  
Balthasar Heesters ◽  
Eric G. Huizinga ◽  
Raoul J. de Groot
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Ligand Specificity ◽  
Receptor Binding Site ◽  
Major Shift ◽  
Murine Coronavirus

scholarly journals The intrinsic factor (IF)-cobalamin receptor binding site is located in the amino-terminal portion of IF

1992 ◽  
Vol 267 (32) ◽  
pp. 22982-22986
Author(s):  
L.H. Tang ◽  
H Chokshi ◽  
C.B. Hu ◽  
M.M. Gordon ◽  
D.H. Alpers
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Intrinsic Factor ◽  
Terminal Portion ◽  
Receptor Binding Site ◽  
Amino Terminal

scholarly journals N-Linked Glycans and K147 Residue on Hemagglutinin Synergize To Elicit Broadly Reactive H1N1 Influenza Virus Antibodies

2019 ◽  
Vol 94 (6) ◽  
Author(s):  
Ying Huang ◽  
Simon O. Owino ◽  
Corey J. Crevar ◽  
Donald M. Carter ◽  
Ted M. Ross
Keyword(s):  
Amino Acids ◽  
Influenza Virus ◽  
Binding Site ◽  
Receptor Binding ◽  
Hemagglutination Inhibition ◽  
Vaccine Development ◽  
H1n1 Influenza ◽  
Influenza Viruses ◽  
Wild Type ◽  
Receptor Binding Site

ABSTRACT Vaccination is the most effective way to prevent influenza virus infections. However, the diversity of antigenically distinct isolates is a challenge for vaccine development. In order to overcome the antigenic variability and improve the protective efficacy of influenza vaccines, our research group has pioneered the development of computationally optimized broadly reactive antigens (COBRA) for hemagglutinin (HA). Two candidate COBRA HA vaccines, P1 and X6, elicited antibodies with differential patterns of hemagglutination inhibition (HAI) activity against a panel of H1N1 influenza viruses. In order to better understand how these HA antigens elicit broadly reactive immune responses, epitopes in the Cb, Sa, or Sb antigenic sites of seasonal-like and pandemic-like wild-type or COBRA HA antigens were exchanged with homologous regions in the COBRA HA proteins to determine which regions and residues were responsible for the elicited antibody profile. Mice were vaccinated with virus-like particles (VLPs) expressing one of the 12 modified HA antigens (designated V1 to V12), COBRA HA antigens, or wild-type HA antigens. The elicited antisera was assessed for hemagglutination inhibition activity against a panel of historical seasonal-like and pandemic-like H1N1 influenza viruses. Primarily, the pattern of glycosylation sites and residues in the Sa antigenic region, around the receptor binding site (RBS), served as signatures for the elicitation of broadly reactive antibodies by these HA immunogens. Mice were vaccinated with VLPs expressing HA antigens that lacked a glycosylation site at residue 144 and a deleted lysine at position 147 residue were more effective at protecting against morbidity and mortality following infection with pandemic-like and seasonal-like H1N1 influenza viruses. IMPORTANCE There is a great need to develop broadly reactive or universal vaccines against influenza viruses. Advanced, next-generation hemagglutinin (HA) head-based vaccines that elicit protective antibodies against H1N1 influenza viruses have been developed. This study focused on understanding the specific amino acids around the receptor binding site (RBS) that were important in elicitation of these broadly reactive antibodies. Specific glycan sites and amino acids located at the tip of the HA molecule enhanced the elicitation of these broadly reactive antibodies. A better understanding of the HA structures around the RBS will lead to more effective HA immunogens.

scholarly journals Evidence for the location of the receptor-binding site of human erythropoietin at the carboxyl-terminal domain

Blood ◽  
1991 ◽  
Vol 77 (6) ◽  
pp. 1203-1210
Author(s):  
MR Fibi ◽  
W Stuber ◽  
P Hintz-Obertreis ◽  
G Luben ◽  
D Krumwieh ◽  
...  
Keyword(s):  
Binding Site ◽  
Receptor Binding ◽  
Terminal Region ◽  
Biologically Active ◽  
Receptor Binding Site ◽  
Specific Antibodies ◽  
Amino Terminal ◽  
Human Erythropoietin ◽  
A Cell ◽  
Carboxyl Terminal

Five different peptides (P1: 84′95; P2: 152′166; P3: 52′63; P4: 7′23; P5: 110′123) homologous to relatively hydrophilic regions of human erythropoietin (huEpo) have been synthesized to identify biologically active domains of the hormone. All peptides were able to induce high titers of peptide-specific antibodies in rabbits. Antisera from rabbits induced by recombinant huEpo (rhuEpo) contained a relatively high amount of antibodies preferentially directed against three peptides (P2, P4, and P5), of which P4 comprised the amino-terminal region, P2 the carboxyl-terminus, and P5 an interior region previously described as the receptor-binding site. The same three peptides were able to induce rhuEpo-specific antibodies, whereas P1 and P3 lacked this activity. Only peptide-P2-induced antisera inhibited the biologic activity of rhuEpo in a cell proliferation assay, indicating that the carboxyl-terminal region of the molecule is essentially involved in the biologic function of rhuEpo.

scholarly journals Amino Acids 270 to 510 of the Severe Acute Respiratory Syndrome Coronavirus Spike Protein Are Required for Interaction with Receptor

2004 ◽  
Vol 78 (9) ◽  
pp. 4552-4560 ◽  
Author(s):  
Gregory J. Babcock ◽  
Diana J. Esshaki ◽  
William D. Thomas ◽  
Donna M. Ambrosino
Keyword(s):  
Amino Acids ◽  
Severe Acute Respiratory Syndrome ◽  
Receptor Binding ◽  
Expression System ◽  
Viral Envelope ◽  
Receptor Binding Site ◽  
Host Cell Membrane ◽  
Viral Attachment ◽  
Amino Terminal ◽  
Mammalian Expression

ABSTRACT A novel coronavirus, severe acute respiratory syndrome coronavirus (SARS-CoV), has recently been identified as the causative agent of severe acute respiratory syndrome (SARS). SARS-CoV appears similar to other coronaviruses in both virion structure and genome organization. It is known for other coronaviruses that the spike (S) glycoprotein is required for both viral attachment to permissive cells and for fusion of the viral envelope with the host cell membrane. Here we describe the construction and expression of a soluble codon-optimized SARS-CoV S glycoprotein comprising the first 1,190 amino acids of the native S glycoprotein (S1190). The codon-optimized and native S glycoproteins exhibit similar molecular weight as determined by Western blot analysis, indicating that synthetic S glycoprotein is modified correctly in a mammalian expression system. S1190 binds to the surface of Vero E6 cells, a cell permissive to infection, as demonstrated by fluorescence-activated cell sorter analysis, suggesting that S1190 maintains the biologic activity present in native S glycoprotein. This interaction is blocked with serum obtained from recovering SARS patients, indicating that the binding is specific. In an effort to map the ligand-binding domain of the SARS-CoV S glycoprotein, carboxy- and amino-terminal truncations of the S1190 glycoprotein were constructed. Amino acids 270 to 510 were the minimal receptor-binding region of the SARS-CoV S glycoprotein as determined by flow cytometry. We speculate that amino acids 1 to 510 of the SARS-CoV S glycoprotein represent a unique domain containing the receptor-binding site (amino acids 270 to 510), analogous to the S1 subunit of other coronavirus S glycoproteins.

scholarly journals Human coronaviruses OC43 and HKU1 bind to 9-O-acetylated sialic acids via a conserved receptor-binding site in spike protein domain A

2019 ◽  
Vol 116 (7) ◽  
pp. 2681-2690 ◽  
Author(s):  
Ruben J. G. Hulswit ◽  
Yifei Lang ◽  
Mark J. G. Bakkers ◽  
Wentao Li ◽  
Zeshi Li ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Binding Site ◽  
Receptor Binding ◽  
Protein S ◽  
Protein Domain ◽  
Spike Protein ◽  
Ligand Docking ◽  
Respiratory Pathogens ◽  
Receptor Binding Site ◽  
Human Coronaviruses

Human betacoronaviruses OC43 and HKU1 are endemic respiratory pathogens and, while related, originated from independent zoonotic introductions. OC43 is in fact a host-range variant of the species Betacoronavirus-1, and more closely related to bovine coronavirus (BCoV)—its presumptive ancestor—and porcine hemagglutinating encephalomyelitis virus (PHEV). The β1-coronaviruses (β1CoVs) and HKU1 employ glycan-based receptors carrying 9-O-acetylated sialic acid (9-O-Ac-Sia). Receptor binding is mediated by spike protein S, the main determinant of coronavirus host specificity. For BCoV, a crystal structure for the receptor-binding domain S1A is available and for HKU1 a cryoelectron microscopy structure of the complete S ectodomain. However, the location of the receptor-binding site (RBS), arguably the single-most important piece of information, is unknown. Here we solved the 3.0-Å crystal structure of PHEV S1A. We then took a comparative structural analysis approach to map the β1CoV S RBS, using the general design of 9-O-Ac-Sia-binding sites as blueprint, backed-up by automated ligand docking, structure-guided mutagenesis of OC43, BCoV, and PHEV S1A, and infectivity assays with BCoV-S–pseudotyped vesicular stomatitis viruses. The RBS is not exclusive to OC43 and related animal viruses, but is apparently conserved and functional also in HKU1 S1A. The binding affinity of the HKU1 S RBS toward short sialoglycans is significantly lower than that of OC43, which we attribute to differences in local architecture and accessibility, and which may be indicative for differences between the two viruses in receptor fine-specificity. Our findings challenge reports that would map the OC43 RBS elsewhere in S1A and that of HKU1 in domain S1B.

Sign in / Sign up

Export Citation Format

Share Document