Cloned (Comment) | Organism |
---|---|
gene luc, DNA and amino acid sequence determination and analysis, sequence comparisons of Photinus pyralis and Photinus scintillans luciferases, the single conservative amino acid change tyrosine to phenylalanine at position 255 accounts for the entire emission color difference, recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) pLysS, recombinant Luc contains the N-terminal peptide GlyProLeuGlySer- | Photinus pyralis |
gene luc, DNA and amino acid sequence determination and analysis, sequence comparisons of Photinus pyralis and Photinus scintillans luciferases, the single conservative amino acid change tyrosine to phenylalanine at position 255 accounts for the entire emission color difference, recombinant expression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) pLysS, recombinant Luc contains the N-terminal peptide GlyProLeuGlySer- | Photinus scintillans |
Protein Variants | Comment | Organism |
---|---|---|
E311Q | site-directed mutagenesis, the mutant shows 65% specific activity compared to the wild-type | Photinus pyralis |
F255Y | site-directed mutagenesis, the single change Y255F is necessary and sufficient for a 13 nm redshift in bioluminescence | Photinus scintillans |
additional information | Photinus scintillans luciferases PsnWT is significantly more resistant to long wavelength emission shifts than Photinus pyralis luciferase PpyWT at pH 7.0 and pH 6.0 | Photinus pyralis |
additional information | Photinus scintillans luciferases PsnWT is significantly more resistant to long wavelength emission shifts than Photinus pyralis luciferase PpyWT at pH 7.0 and pH 6.0 | Photinus scintillans |
N229T | site-directed mutagenesis, the mutant shows 63% specific activity compared to the wild-type | Photinus pyralis |
R213K/T214N | site-directed mutagenesis, residues K213 and/or N214 are largely responsible for the 1.55fold increase in specific activity of the variant | Photinus pyralis |
R218Q | site-directed mutagenesis, the mutant shows 125% specific activity compared to the wild-type | Photinus pyralis |
R337Q | site-directed mutagenesis, the mutant shows 26% specific activity compared to the wild-type | Photinus pyralis |
S284T | site-directed mutagenesis, a red-emitting mutant variant, the mutant shows 25% specific activity compared to the wild-type | Photinus pyralis |
Y255F | site-directed mutagenesis, the mutant shows 71% specific activity compared to the wild-type | Photinus pyralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | the Km value for the reaction intermediate D-firefly luciferin-AMP is 0.14 mM | Photinus pyralis | |
additional information | - |
additional information | the Km value for the reaction intermediate D-firefly luciferin-AMP is 0.24 mM | Photinus scintillans | |
0.014 | - |
D-firefly luciferin | pH 7.8, 22°C, recombinant mutant Y255F | Photinus pyralis | |
0.015 | - |
D-firefly luciferin | pH 7.8, 22°C, recombinant mutant F255Y | Photinus scintillans | |
0.015 | - |
D-firefly luciferin | pH 7.8, 22°C, recombinant wild-type enzyme | Photinus pyralis | |
0.019 | - |
D-firefly luciferin | pH 7.8, 22°C, recombinant wild-type enzyme | Photinus scintillans | |
0.024 | - |
D-firefly luciferin | pH 7.8, 22°C, recombinant mutant R213K/T214N | Photinus pyralis | |
0.08 | - |
ATP | pH 7.8, 22°C, recombinant mutant Y255F | Photinus pyralis | |
0.086 | - |
ATP | pH 7.8, 22°C, recombinant wild-type enzyme | Photinus pyralis | |
0.126 | - |
ATP | pH 7.8, 22°C, recombinant mutant R213K/T214N | Photinus pyralis | |
0.135 | - |
ATP | pH 7.8, 22°C, recombinant wild-type enzyme | Photinus scintillans | |
0.159 | - |
ATP | pH 7.8, 22°C, recombinant mutant F255Y | Photinus scintillans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-firefly luciferin + O2 + ATP | Photinus pyralis | - |
firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? | |
D-firefly luciferin + O2 + ATP | Photinus scintillans | - |
firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | P08659 | - |
- |
Photinus scintillans | A0A1B3TNR9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) pLysS by glutathione affinity chromatography | Photinus pyralis |
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) pLysS by glutathione affinity chromatography | Photinus scintillans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-firefly luciferin + O2 + ATP | - |
Photinus pyralis | firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? | |
D-firefly luciferin + O2 + ATP | - |
Photinus scintillans | firefly oxyluciferin + CO2 + AMP + diphosphate + hv | - |
? | |
additional information | analysis of reaction parameters, detailed overview | Photinus pyralis | ? | - |
? | |
additional information | analysis of reaction parameters, detailed overview | Photinus scintillans | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
firefly luciferase | - |
Photinus pyralis |
Luc | - |
Photinus pyralis |
Luc | - |
Photinus scintillans |
luciferase | - |
Photinus pyralis |
luciferase | - |
Photinus scintillans |
orange light-producing luciferase | - |
Photinus scintillans |
PpyWT | - |
Photinus pyralis |
PsntWT | - |
Photinus scintillans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
- |
Photinus pyralis |
22 | - |
- |
Photinus scintillans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 240 min | Photinus scintillans |
37 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 47 min | Photinus pyralis |
42 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 50 s | Photinus pyralis |
42 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 61 s | Photinus scintillans |
45 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 25 s | Photinus pyralis |
45 | - |
thermal inactivation of purified recombinant wild-type enzyme at pH 7.4 after 40 s | Photinus scintillans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.8 | - |
- |
Photinus pyralis |
7.8 | - |
- |
Photinus scintillans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Photinus pyralis | |
ATP | - |
Photinus scintillans |
General Information | Comment | Organism |
---|---|---|
malfunction | the single conservative amino acid change tyrosine to phenylalanine at position 255 accounts for the entire emission color difference | Photinus pyralis |
malfunction | the single conservative amino acid change tyrosine to phenylalanine at position 255 accounts for the entire emission color difference | Photinus scintillans |
physiological function | beetle bioluminescence is the result of two half-reactions catalyzed by a luciferase (Luc) in which the substrate D-firefly luciferin is converted into the corresponding adenylate D-firefly luciferin-AMP, followed by a multistep oxidative process that produces light via a single electron transfer process. Among the Lampyridae, one of the three families of bioluminescent beetles, light emission typically ranges from green to yellow-green in color having emission maxima (lambdamax) of about 548-568 nm | Photinus pyralis |
physiological function | beetle bioluminescence is the result of two half-reactions catalyzed by a luciferase (Luc) in which the substrate D-firefly luciferin is converted into the corresponding adenylate D-firefly luciferin-AMP, followed by a multistep oxidative process that produces light via a single electron transfer process. Among the Lampyridae, one of the three families of bioluminescent beetles, light emission typically ranges from green to yellow-green in color having emission maxima (lambdamax) of about 548-568 nm | Photinus scintillans |