Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as a His-tagged fusion protein | Photinus pyralis |
Protein Variants | Comment | Organism |
---|---|---|
D474K | D476N mutation does not have any significant effect | Photinus pyralis |
D476N | D474K mutation destabilizes the protein. Flexibility analysis using dynamic quenching and limited proteolysis demonstrates that D474K mutation is much more flexible than wild-type | Photinus pyralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
D-luciferin | mutant D476N, pH 7, 25°C | Photinus pyralis | |
0.015 | - |
D-luciferin | wild-type, pH 8, 25°C | Photinus pyralis | |
0.016 | - |
D-luciferin | mutant D474K, pH 8.5, 25°C | Photinus pyralis | |
0.1 | - |
ATP | mutant D476N, pH 7, 25°C | Photinus pyralis | |
0.1 | - |
ATP | wild-type, pH 8, 25°C | Photinus pyralis | |
0.15 | - |
ATP | mutant D474K, pH 8.5, 25°C | Photinus pyralis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
using Ni-NTA chromatography | Photinus pyralis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
relative specific activity: wild-type 100%, mutant D474K 55%, mutant D476N 181% | Photinus pyralis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-luciferin + ATP + O2 | - |
Photinus pyralis | oxidized D-luciferin + AMP + CO2 + diphosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
luciferin | - |
Photinus pyralis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Photinus pyralis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | 45 | when native and mutants are incubated 5 min at temperatures ranging from 15 to 45°C, D474K mutant is inactivated faster than native and D476N at all temperatures | Photinus pyralis |
32 | - |
after 20 min at 32°C wild-type and mutant D476N retain 24% original activity whereas mutant D474K shows only 2% remaining activity | Photinus pyralis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
mutant D476N | Photinus pyralis |
8 | - |
wild-type | Photinus pyralis |
8.5 | - |
mutant D474K | Photinus pyralis |