Cloned (Comment) | Organism |
---|---|
expressed in Drosophila melanogaster Schneider S2 cells | influenza A virus H1N1 |
Crystallization (Comment) | Organism |
---|---|
mutant enzymes in complex with oseltamivir carboxylate, hanging drop vapor diffusion method, mutant I223V: 0.1 M HEPES pH 6.75, 7% (w/v) PEG 8000, mutant S247N: 0.1 M HEPES pH 6.7, 8.5% (w/v) PEG 8000, mutant H275Y: 0.1 M HEPES pH 7.0, 8% (w/v) PEG 8000, mutant I223V/H275Y: 0.1 M HEPES pH 7.0, 7% (w/v) PEG 8000, and mutant S247N/H275Y: 0.1 M HEPES pH 7.5, 5% (w/v) PEG 8000 | influenza A virus H1N1 |
Protein Variants | Comment | Organism |
---|---|---|
H275Y | the major oseltamivir resistance mutation H275Y causes a significant decrease in the enzyme's ability to bind oseltamivir | influenza A virus H1N1 |
H275Y/I223V | the mutations result in extreme impairment of oseltamivir's inhibition potency | influenza A virus H1N1 |
H275Y/S247N | the mutations result in extreme impairment of oseltamivir's inhibition potency | influenza A virus H1N1 |
I223V | the substitution alone confers only a moderate reduction in oseltamivir affinity | influenza A virus H1N1 |
S247N | the substitution alone confers only a moderate reduction in oseltamivir affinity | influenza A virus H1N1 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
oseltamivir | - |
influenza A virus H1N1 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.1 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | wild type enzyme, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
1.1 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
1.1 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
1.2 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
1.3 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
3.3 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
influenza A virus H1N1 | C3W6G3 | A/California/07/2009 | - |
Purification (Comment) | Organism |
---|---|
Strep-Tactin resin column chromatography | influenza A virus H1N1 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid + H2O | - |
influenza A virus H1N1 | 4-methylumbelliferone + alpha-D-N-acetylneuraminic acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
neuraminidase | - |
influenza A virus H1N1 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.054 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.1 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.17 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.6 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.9 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | wild type enzyme, at 37°C, pH not specified in the publication | influenza A virus H1N1 |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000024 | - |
oseltamivir | wild type enzyme, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.00045 | - |
oseltamivir | mutant enzyme S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.0007 | - |
oseltamivir | mutant enzyme I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.027 | - |
oseltamivir | mutant enzyme H275Y, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.094 | - |
oseltamivir | mutant enzyme H275Y/I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.22 | - |
oseltamivir | mutant enzyme H275Y/S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.035 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y/I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.048 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme H275Y, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.16 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme S247N, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.51 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | mutant enzyme I223V, at 37°C, pH not specified in the publication | influenza A virus H1N1 | |
0.82 | - |
4-methylumbelliferyl alpha-D-N-acetylneuraminic acid | wild type enzyme, at 37°C, pH not specified in the publication | influenza A virus H1N1 |