Application | Comment | Organism |
---|---|---|
medicine | the enzyme is a potential therapeutic target for neurodegenerative and neurologic disorders | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli and in HEK293T cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
crystals are obtained by hanging-drop vapor diffusion at 20°C. Crystal structures of the complex of the full-length enzyme with the substrate L-kynurenine and in complex with L-kynurenine and Ro 61-8048 at a resolution of 1.85 and 2.34 A, respectively. The crystals of the enzyme-L-kynurenine complex belong to the space group P2(1) with 1 enzyme molecule in the asymmetric unit. The crystal structure of the enzymeL-kynurenine-Ro61-8048 complex belongs to the space group P2(1)2(1)2(1) with 1 pfKMO molecule in the asymmetric unit. The crystal structure of the SeMet enzyme derivative belongs to the space group P2(1) with 2 enzyme molecules in the asymmetric unit | Pseudomonas fluorescens |
Protein Variants | Comment | Organism |
---|---|---|
E366Q | 2% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
E372A | about 15% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
E372Q | about 5% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
M367A | 2% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
M367L | 13% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
M373A | no activity detected | Pseudomonas fluorescens |
M373L | about 60% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
additional information | the enzyme is systematically truncated according to the sequence alignments and the predicted secondary structures. For the truncations 1-377, 1-379, and 1-394, which have 1 or 2 more predicted alpha-helices than that of the 1-372/374, no or weak protein expression is detected. Although for the truncation 1-430 that has the C-terminal hydrophobic tail removed, no enzymatic activities can be detected but the protein expression is normal. The results indicate that the C-terminal portion is important for both the folding and the enzymatic activity | Homo sapiens |
N363A | 28% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
N363D | no activity detected | Homo sapiens |
N369A | about 65% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
N369D | no activity detected | Pseudomonas fluorescens |
N465A | about 80% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
Q424A | mutation does not greatly affect enzyme activity. Ro 61-8048 shows no inhibition to the pfKMO mutant enzyme | Pseudomonas fluorescens |
R84A | no activity detected | Pseudomonas fluorescens |
R85A | no activity detected | Homo sapiens |
R85K | 1% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
Y398A | 1% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
Y398F | 1% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
Y404A | no activity detected | Pseudomonas fluorescens |
Y404F | about 40% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
Y98A | no activity detected | Pseudomonas fluorescens |
Y98F | about 1% of the enzyme activity compared with that of the wild-type enzyme | Pseudomonas fluorescens |
Y99A | no activity detected | Homo sapiens |
Y99F | 7% of the enzyme activity compared with that of the wild-type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ro 61-8048 | allosteric | Homo sapiens | |
Ro 61-8048 | noncompetitive. Theinhibitor is bound in the tunnel at the interface where the N- and C-terminal domains associate | Pseudomonas fluorescens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
60386 | - |
calculated from sequence | Homo sapiens |
62136 | - |
native mass spectrum analysis | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | O15229 | - |
- |
Pseudomonas fluorescens | Q84HF5 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | glycosylation has only a marginal effect on enzyme activity | Homo sapiens |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas fluorescens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-kynurenine + NADPH + H+ + O2 | - |
Pseudomonas fluorescens | 3-hydroxy-L-kynurenine + NADP+ + H2O | - |
? | |
L-kynurenine + NADPH + H+ + O2 | - |
Homo sapiens | 3-hydroxy-L-kynurenine + NADP+ + H2O | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Pseudomonas fluorescens |
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pseudomonas fluorescens |
8 | - |
assay at | Homo sapiens |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0135 | - |
Ro 61-8048 | pH and temperature not specified in the publication | Pseudomonas fluorescens |
General Information | Comment | Organism |
---|---|---|
drug target | the enzyme is a potential therapeutic target for neurodegenerative and neurologic disorders | Homo sapiens |