Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
molecular modeling of enzyme and enzyme-spermine complex. Spermine oxidase binds spermine in a similar conformation as that observed in the yeast polyamine oxidase FMS1-spermine complex, with a major role for residues H82 and K367 in substrate binding and catalysis. The enzymesubstrate complex shows an active site pocket with highly polar characteristics | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
H82E | no catalytic activity | Mus musculus |
H82E | site-directed mutagenesis, inactive mutant | Mus musculus |
H82Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Mus musculus |
H82Q | about 300fold decrease in catalytic efficiency | Mus musculus |
K367M | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Mus musculus |
K367M | about 3fold decrease in catalytic efficiency | Mus musculus |
additional information | wild-type and mutant proteins share essentially the same structural properties. The pH-dependency of the catalytic activity of H82Q is essentially unchanged with respect to the wild-type, while a slight shift toward higher pH values is observed for the K367M mutant | Mus musculus |
T428Y | no catalytic activity | Mus musculus |
T528Y | site-directed mutagenesis, inactive mutant | Mus musculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
N1,N4-bis(2,3-butadienyl)-1,4-butanediamine | i.e. MDL72527, inhibits FAD-containing polyamine oxidases | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
spermine | mutant K367M, pH 8.5, 25°C | Mus musculus | |
0.021 | - |
spermine | pH 8.5, 25°C, recombinant mutant K367M | Mus musculus | |
0.242 | - |
spermine | wild-type, pH 8.5, 25°C | Mus musculus | |
0.242 | - |
spermine | pH 8.5, 25°C, recombinant wild-type enzyme | Mus musculus | |
0.53 | - |
spermine | mutant H82Q, pH 8.5, 25°C | Mus musculus | |
0.53 | - |
spermine | pH 8.5, 25°C, recombinant mutant H82Q | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
spermine + O2 + H2O | Mus musculus | - |
spermidine + 3-aminopropanal + H2O2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
- |
- |
Mus musculus | Q99K82 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
spermine + O2 + H2O | - |
Mus musculus | spermidine + 3-aminopropanal + H2O2 | - |
? | |
spermine + O2 + H2O | SMO oxidizes the carbon on the exo side of the N5-nitrogen of SPM producing spermidine, 3-aminopropanal, and H2O2 | Mus musculus | spermidine + 3-aminopropanal + H2O2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
SMO | - |
Mus musculus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.024 | - |
spermine | mutant H82Q, pH 8.5, 25°C | Mus musculus | |
0.024 | - |
spermine | pH 8.5, 25°C, recombinant mutant H82Q | Mus musculus | |
0.1 | - |
spermine | mutant K367M, pH 8.5, 25°C | Mus musculus | |
0.1 | - |
spermine | pH 8.5, 25°C, recombinant mutant K367M | Mus musculus | |
3.57 | - |
spermine | wild-type, pH 8.5, 25°C | Mus musculus | |
3.57 | - |
spermine | pH 8.5, 25°C, recombinant wild-type enzyme | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Mus musculus |
8.5 | - |
wild-type and mutant H82Q | Mus musculus |
9 | 9.5 | mutant K367M | Mus musculus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 8.5 | recombinant wild-type MmSMO enzymatic activity increases in the pH range, reaching a maximum at pH 8.5 and decreasing for higher pH values | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on | Mus musculus |
General Information | Comment | Organism |
---|---|---|
additional information | substrate binding and catalytic mechanism, spermine docking into the active site, CD spectroscopy, structure modelling, overview | Mus musculus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00000005 | - |
spermine | pH 8.5, 25°C, recombinant mutant H82Q | Mus musculus | |
0.000005 | - |
spermine | pH 8.5, 25°C, recombinant mutant K367M | Mus musculus | |
0.000015 | - |
spermine | pH 8.5, 25°C, recombinant wild-type enzyme | Mus musculus | |
0.00005 | - |
spermine | mutant H82Q, pH 8.5, 25°C | Mus musculus | |
0.0015 | - |
spermine | wild-type, pH 8.5, 25°C | Mus musculus | |
0.005 | - |
spermine | mutant K367M, pH 8.5, 25°C | Mus musculus |