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Literature summary for 1.13.12.7 extracted from
- Design and characterization of novel trypsin-resistant firefly luciferases by site-directed mutagenesis (2009), Protein Eng. Des. Sel., 22, 655-663.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli BL21 and DH5alpha, transfection in CHO-cells | Photinus pyralis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| Q338P | about 50% decrease in specific activity compared to wild-type luciferase | Photinus pyralis |
| R213E | same activity like wild-type luciferase | Photinus pyralis |
| R213M | 3fold activity compared to wild-type luciferase | Photinus pyralis |
| R337Q | only 60% relative specific activity compared to wild-type luciferase, higher light stability, most stable luciferase mutant against trypsin hydrolysis at 23 and 37°C | Photinus pyralis |
General Stability
| General Stability | Organism |
|---|---|
| decrease of remaining activity after trypsin hydrolysis at 23 and 37°C, mutants R213M and R337Q are more stable than the wild-type luciferase | Photinus pyralis |
| mutant R337Q still 26% remaining activity after 15 min trypsin hydrolysis at 37°C | Photinus pyralis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.01 | - |
Luciferin | mutant R213M | Photinus pyralis |  |
| 0.012 | - |
Luciferin | wild-type luciferase | Photinus pyralis | |
| 0.013 | - |
Luciferin | mutant Q338P | Photinus pyralis | |
| 0.014 | - |
Luciferin | mutant R213E | Photinus pyralis | |
| 0.018 | - |
Luciferin | mutant R337Q | Photinus pyralis | |
| 0.133 | - |
ATP | wild-type luciferase | Photinus pyralis | |
| 0.15 | - |
ATP | mutant R213M | Photinus pyralis | |
| 0.2 | - |
ATP | mutant R213E | Photinus pyralis | |
| 0.35 | - |
ATP | mutant Q338P | Photinus pyralis | |
| 0.5 | - |
ATP | mutant R337Q | Photinus pyralis | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| intracellular | - |
Photinus pyralis | 5622 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | required | Photinus pyralis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 62000 | - |
SDS-PAGE | Photinus pyralis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Photinus pyralis | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-NTA spin column | Photinus pyralis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| luciferin + ATP + O2 | assay at 25°C, pH 7.8 | Photinus pyralis | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| firefly luciferase | - |
Photinus pyralis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
- |
Photinus pyralis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.3 | - |
mutant R337Q | Photinus pyralis |
| 7.8 | - |
assay at | Photinus pyralis |
| 8 | - |
- |
Photinus pyralis |
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Release 2023.1 (January 2023)
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