Application | Comment | Organism |
---|---|---|
drug development | broad-spectrum antivirals that target orthomyxo- and coronavirus sialate-O-acetylesterases | bovine coronavirus |
molecular biology | origin and evolution of viral hemagglutinin-esterases | bovine coronavirus |
Cloned (Comment) | Organism |
---|---|
sequence encoding ectodomain (residues 19-388) in plasmid pCD5-BCoVHE-T-Fc for expression with N-terminal CD5 signal peptide and C-terminal thrombin cleavage site followed by human IgG1 Fc domain in N-acetylglucosaminyltransferase-deficient HEK293S cells | bovine coronavirus |
Crystallization (Comment) | Organism |
---|---|
apo-enzyme (PDB: 3CL4) or mutant S40A in complex with 5-N-acetyl-4,9-di-O-acetylneuraminic acid alpha-methylglycoside (PDB: 3CL5), hanging drop vapour diffusion, 18°C, 2 weeks, precipitant: 16% polyethylene glycol 8000 and 20% glycerol or 10% polyethylene glycol 3350, hexagonal bipyramide crystals: space groups: P6(5)22, unit cell parameter: a, b: 88.8-89.3, c: 280.4-282.4, soaking with 7 mM 5-N-acetyl-4,9-di-O-acetylneuraminic acid alpha-methylglycoside, molecular replacement using PDB: 1FLC (for apo-enzyme) or wild-type structure (for S40A mutant) as template, three domains: receptor-binding domain (R), acetylase domain (E, strictly conserved), and membrane-proximal domain (MP), homodimer of 2fold crystallographic symmetry with 2 major contact regions (CR1: bridges R domains, CR2: involves MP), disorder of residues 377-388, E-domain: SGNH-hydrolase fold, active site with catalytic triad (Ser40, His329, Asp326) and oxyanion hole (Asn104, Ser40, Gly75), highly variable surface loop (residues 47-54) is site of antigenic variation, R-domain: different from those of hemagglutinin-esterase fusion (HEF) and agglutinin (HA) (plasticity), binding of ligand in opposite orientation involving residues Leu212, Asn214, Ser213, Tyr184, Phe211, Leu266, Leu267 (hydrophobic pocket), coordination of potassium and water by Asp220, Ser221, Gln222, Ser263, Glu265, Leu267 | bovine coronavirus |
Protein Variants | Comment | Organism |
---|---|---|
F211A | abrogated ligand recognition and binding | bovine coronavirus |
L266A | abrogated ligand recognition and binding | bovine coronavirus |
L267A | abrogated ligand recognition and binding | bovine coronavirus |
S40A | catalytically inactive, active site residue, retained lectin activity | bovine coronavirus |
Y184A | decreased ligand binding affinity | bovine coronavirus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | bovine coronavirus | purified enzyme destroys BCoV receptors on rat erythrocytes and Madin-Darby bovine kidney (MDBK) cells | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
bovine coronavirus | P15776 | BcoV, strain Mebus | - |
Purification (Comment) | Organism |
---|---|
from HEK293S cells by protein A-affinity chromatography followed by on-bead thrombin digestion | bovine coronavirus |
Source Tissue | Comment | Organism | Textmining |
---|
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-N-acetyl-4,9-di-O-acetylneuraminic acid alpha-methylglycoside + H2O | sialate-9-O-acetylesterase activity | bovine coronavirus | 4,9-di-O-acetylneuraminic acid alpha-methylglycoside + acetate | analysis by gas chromatography/mass spectrometry | ? | |
additional information | purified enzyme destroys BCoV receptors on rat erythrocytes and Madin-Darby bovine kidney (MDBK) cells | bovine coronavirus | ? | - |
? | |
p-nitrophenyl acetate + H2O | - |
bovine coronavirus | p-nitrophenol + acetate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | crystal structure, disulphide-linkage through Cys385, 2fold crystallographic symmetry | bovine coronavirus |
Synonyms | Comment | Organism |
---|---|---|
BCoVHE | - |
bovine coronavirus |
HE | - |
bovine coronavirus |
hemagglutinin-esterase | - |
bovine coronavirus |