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Literature summary for 3.1.1.53 extracted from

  • Zeng, Q.; Langereis, M.A.; van Vliet, A.L.; Huizinga, E.G.; de Groot, R.J.
    Structure of coronavirus hemagglutinin-esterase offers insight into corona and influenza virus evolution (2008), Proc. Natl. Acad. Sci. USA, 105, 9065-9069.
    View publication on PubMedView publication on EuropePMC

Application

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(Commentary)

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 (Commentary)

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

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/ Products (Substrates)

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 ?
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?

Organism

Organism UniProt Comment Textmining
bovine coronavirus P15776 BcoV, strain Mebus
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Purification (Commentary)

Purification (Comment) Organism
from HEK293S cells by protein A-affinity chromatography followed by on-bead thrombin digestion bovine coronavirus

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

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
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bovine coronavirus p-nitrophenol + acetate
-
?

Subunits

Subunits Comment Organism
dimer crystal structure, disulphide-linkage through Cys385, 2fold crystallographic symmetry bovine coronavirus

Synonyms

Synonyms Comment Organism
BCoVHE
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bovine coronavirus
HE
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bovine coronavirus
hemagglutinin-esterase
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bovine coronavirus