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Literature summary for 3.2.1.18 extracted from

  • Albohy, A.; Li, M.D.; Zheng, R.B.; Zou, C.; Cairo, C.W.
    Insight into substrate recognition and catalysis by the human neuraminidase 3 (NEU3) through molecular modeling and site-directed mutagenesis (2010), Glycobiology, 20, 1127-1138.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant A160G, E51S, H277F, R48N, R48S, M87G, D50S, and Y370F enzymes as MBP-fusion proteins in Escherichia coli strain TB1 Homo sapiens

Protein Variants

Protein Variants Comment Organism
A160G site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme Homo sapiens
A160G/M87G/I105G site-directed mutagenesis, inactive mutant Homo sapiens
D50S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
E113A site-directed mutagenesis, inactive mutant Homo sapiens
E225S site-directed mutagenesis, almost inactive mutant Homo sapiens
E51D site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
E51S site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Homo sapiens
G162A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
G419STOP site-directed mutagenesis, the C-terminal 10-residue truncation leads to an increase in activity compared to the wild-type enzyme Homo sapiens
H277F site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Homo sapiens
I105G site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
I407STOP site-directed mutagenesis, almost inactive mutant Homo sapiens
L408STOP site-directed mutagenesis, the large deletion leads to inactivation of the mutant Homo sapiens
M87G site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
M87G/I105G site-directed mutagenesis, inactive mutant Homo sapiens
additional information mutations of residues expected to interact directly with the sialic acid N5-acetyl (A160, M87, I105) and C7-C9 glycerol side-chain (E113, Y179, Y181) reduce enzymatic activity. Truncations at the N- or C-terminus of more than 10 residues abolish enzyme activity Homo sapiens
N88D site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
R114A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
R114Q site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
R245A site-directed mutagenesis, inactive mutant Homo sapiens
R25H site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
R340A site-directed mutagenesis, almost inactive mutant Homo sapiens
R45V site-directed mutagenesis, inactive mutant Homo sapiens
T403STOP site-directed mutagenesis, inactive mutant Homo sapiens
V107M site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Homo sapiens
Y179F site-directed mutagenesis, inactive mutant Homo sapiens
Y181F site-directed mutagenesis, inactive mutant Homo sapiens
Y370C site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Homo sapiens
Y370F site-directed mutagenesis, inactive mutant Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
2-deoxy-2,3-dehydro-N-acetylneuraminic acid
-
Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.018
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant H277F, pH 5.0, 37°C Homo sapiens
0.021
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant A160G, pH 5.0, 37°C Homo sapiens
0.032
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant R48S, pH 5.0, 37°C Homo sapiens
0.033
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant R48N, pH 5.0, 37°C Homo sapiens
0.038
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant E51S, pH 5.0, 37°C Homo sapiens
0.045
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant wild-type enzyme, pH 5.0, 37°C Homo sapiens
0.14
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant M87G, pH 5.0, 37°C Homo sapiens

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane associated, NEU3 Homo sapiens 16020
-

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9UQ49
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes as MBP-fusion proteins from Escherichia coli strain TB1 by amylose affinity chromatography, the tag is cleaved off by Factor Xa Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
colominic acid + H2O = sialic acid + lactose catalytic mechanism, overview Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid + H2O
-
Homo sapiens 4-methylumbelliferol + alpha-D-N-acetylneuraminic acid
-
?
additional information the key catalytic residues of the enzyme consist of the general acid-base D50 and the nucleophilic Y370-E225 pair, molecular modeling to predict residues involved in the recognition and hydrolysis of glycolipid substrates, homology modeling and molecular docking of NEU3, overview Homo sapiens ?
-
?

Synonyms

Synonyms Comment Organism
exo-sialidase
-
Homo sapiens
Neu3
-
Homo sapiens
neuraminidase 3
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant M87G, pH 5.0, 37°C Homo sapiens
23
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant wild-type enzyme, pH 5.0, 37°C Homo sapiens
27
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutants A160G and H277F, pH 5.0, 37°C Homo sapiens
32
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant R48N, pH 5.0, 37°C Homo sapiens
44
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant R48S, pH 5.0, 37°C Homo sapiens
51
-
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid recombinant mutant E51S, pH 5.0, 37°C Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
assay at Homo sapiens

General Information

General Information Comment Organism
malfunction mutations of residues expected to interact directly with the sialic acid N5-acetyl (A160, M87, I105) and C7-C9 glycerol side-chain (E113, Y179, Y181) reduce enzymatic activity. Truncations at the N- or C-terminus of more than 10 residues abolish enzyme activity Homo sapiens