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 | 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 | Comment | Organism | Structure |
---|---|---|---|
2-deoxy-2,3-dehydro-N-acetylneuraminic acid | - |
Homo sapiens |
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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | associated, NEU3 | Homo sapiens | 16020 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9UQ49 | - |
- |
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 | Comment | Organism | Reaction ID |
---|---|---|---|
colominic acid + H2O = sialic acid + lactose | catalytic mechanism, overview | Homo sapiens |
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 | Comment | Organism |
---|---|---|
exo-sialidase | - |
Homo sapiens |
Neu3 | - |
Homo sapiens |
neuraminidase 3 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
assay at | Homo sapiens |
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 |