Cloned (Comment) | Organism |
---|---|
cDNA of mAOX3 cloned from mouse CD1 liver into pMMA1 plasmid and expression as an N-terminal fusion protein with a His6 tag, coexpression with mMCSF, heterologous expression in Escherichia coli | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
vast majority of the crystallization trials performed using the recombinant protein, usable data set of crystals of native mAOX3 with a resolution of 2.9 A | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
benzaldehyde | apparent Km-value of native mAOX3, pH 7.4, 25°C | Mus musculus | |
0.02 | - |
benzaldehyde | apparent Km-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
0.026 | - |
Butanal | apparent Km-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
0.029 | - |
Butanal | apparent Km-value of native mAOX3, pH 7.4, 25°C | Mus musculus | |
0.097 | - |
2-hydroxypyrimidine | apparent Km-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
0.173 | - |
2-hydroxypyrimidine | apparent Km-value of native mAOX3, pH 7.4, 25°C | Mus musculus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
150000 | - |
SDS-PAGE, native and recombinant protein, degradation products of 130000, 80000, 70000, and 55000 Da in case of purified recombinant mAOX3 | Mus musculus |
300000 | - |
gel filtration, native and recombinant mAOX3 | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant mAOX3 purified using sequential Ni-NTA chromatography and size exclusion chromatography, a chemical sulfuration step performed to further increase the activity of the enzyme 1.4fold and after coexpression with mMCSF and chemical sulfuration, 30% of recombinant mAOX3 exists in the catalytically active form, native mAOX3 purified by ammonium sulfate precipitation with 50% saturation, benzamidine Sepharose chromatography and a linear NaCl gradient on a 5/5 FPLC Mono Q column | Mus musculus |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Mus musculus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxypyrimidine + H2O + ferricyanide + O2 | - |
Mus musculus | ? | - |
? | |
benzaldehyde + 2 ferricyanide + H2O | - |
Mus musculus | benzoate + 2 ferrocyanide + 2 H+ | - |
? | |
butanal + 2 ferricyanide + H2O | - |
Mus musculus | butanoate + 2 ferrocyanide + 2 H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 150000, gel filtration | Mus musculus |
Synonyms | Comment | Organism |
---|---|---|
mAOX3 | - |
Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.733 | - |
benzaldehyde | apparent Kcat-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
2.167 | - |
benzaldehyde | apparent Kcat-value of native mAOX3, pH 7.4, 25°C | Mus musculus | |
2.333 | - |
Butanal | apparent Kcat-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
6.4 | - |
Butanal | apparent Kcat-value of native mAOX3, pH 7.4, 25°C | Mus musculus | |
6.88 | - |
2-hydroxypyrimidine | apparent Kcat-value of recombinant mAOX3, pH 7.4, 25°C | Mus musculus | |
21.32 | - |
2-hydroxypyrimidine | apparent Kcat-value of native mAOX3, pH 7.4, 25°C | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Mus musculus | |
molybdenum cofactor | i.e. MoCo | Mus musculus | |
[2Fe-2S]-center | two distinct ones | Mus musculus |
General Information | Comment | Organism |
---|---|---|
physiological function | AOX3 is an enzyme of well known importance in drug metabolism and therefore of increasing importance in recent drug design programs | Mus musculus |