Cloned (Comment) | Organism |
---|---|
wild-type and mutant enzymes expressed as glutathione-S-transferase-fusion proteins containing the additional N-terminal peptide Gly-Pro-Leu-Gly-SER, which remains after PreScission protease cleavage from GST, expression in Escherichia coli | Photinus pyralis |
Protein Variants | Comment | Organism |
---|---|---|
G446I | turnover number is 8.4fold lower compared to wild-type value, KM-value for D-luciferin is 1.5fold lower compared to wild-type value, KM-value for MgATP2- is 2.2fold lower compared to wild-type value, the bioluminescence emission maximum is 554 nm, compared to 558 nm for the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 5.1fold lower compared to wild-type value | Photinus pyralis |
K443A | turnover number is 2655 fold lower compared to wild-type value, KM-value for D-luciferin is 6.5fold lower compared to wild-type value, KM-value for MgATP2- is 3.3fold lower compared to wild-type value, the bioluminescence emission maximum is identical to the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 181fold lower compared to wild-type value | Photinus pyralis |
K443A/K529A | turnover number is 1063700fold lower compared to wild-type value, KM-value for D-luciferin is 4.5fold higher compared to wild-type value, KM-value for MgATP2- is 3.5fold higher compared to wild-type value, the bioluminescence emission maximum is 596 nm, compared to 558 nm for the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 858fold lower compared to wild-type value | Photinus pyralis |
K445Q | turnover number is 1.4fold higher compared to wild-type value, KM-value for D-luciferin is 1.7fold lower compared to wild-type value, KM-value for MgATP2- is 2.3fold lower compared to wild-type value, the bioluminescence emission maximum is 556 nm, compared to 558 nm for the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 1.7fold higher compared to wild-type value | Photinus pyralis |
K529A | turnover number is 668fold lower compared to wild-type value, KM-value for D-luciferin is 15.3fold higher compared to wild-type value, KM-value for MgATP2- is 7.5fold higher compared to wild-type value, the bioluminescence emission maximum is 562 nm, compared to 558 nm for the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 3.5fold lower compared to wild-type value | Photinus pyralis |
Q448A | turnover number is 1.9fold lower compared to wild-type value, KM-value for D-luciferin is 2.5fold lower compared to wild-type value, KM-value for MgATP2- is 2.1fold higher compared to wild-type value, the bioluminescence emission maximum is 557 nm, compared to 558 nm for the wild-type value. The ratio of turnover-number to KM-value for D-luciferyl-O-adenosine monophosphate is 1.1fold lower compared to wild-type value | Photinus pyralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00038 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K443A | Photinus pyralis | |
0.00055 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K443A/K529A | Photinus pyralis | |
0.0015 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme G446I | Photinus pyralis | |
0.0022 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme Q448A | Photinus pyralis | |
0.0023 | - |
D-luciferin | mutant enzyme K443A | Photinus pyralis | |
0.0039 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K445Q | Photinus pyralis | |
0.0047 | - |
D-luciferyl-O-adenosine monophosphate | wild-type enzyme | Photinus pyralis | |
0.006 | - |
D-luciferin | mutant enzyme G446I | Photinus pyralis | |
0.006 | - |
D-luciferin | mutant enzyme Q448A | Photinus pyralis | |
0.0089 | - |
D-luciferin | mutant enzyme K445Q | Photinus pyralis | |
0.0126 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K529A | Photinus pyralis | |
0.015 | - |
D-luciferin | wild-type enzyme | Photinus pyralis | |
0.0486 | - |
MgATP2- | mutant enzyme K443A | Photinus pyralis | |
0.067 | - |
D-luciferin | mutant enzyme K443A/K529A | Photinus pyralis | |
0.069 | - |
MgATP2- | mutant enzyme K445Q | Photinus pyralis | |
0.073 | - |
MgATP2- | mutant enzyme G446I | Photinus pyralis | |
0.16 | - |
MgATP2- | wild-type enzyme | Photinus pyralis | |
0.23 | - |
D-luciferin | mutant enzyme K529A | Photinus pyralis | |
0.341 | - |
MgATP2- | mutant enzyme Q448A | Photinus pyralis | |
0.56 | - |
MgATP2- | mutant enzyme K443A/K529A | Photinus pyralis | |
1.2 | - |
MgATP2- | mutant enzyme K529A | Photinus pyralis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Photinus pyralis | P08659 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-luciferin + O2 + ATP | - |
Photinus pyralis | oxidized luciferin + CO2 + H2O + AMP + diphosphate + hv | - |
? | |
D-luciferyl-O-adenosine monophosphate + ? | - |
Photinus pyralis | ? | - |
? | |
luciferin + MgATP2- | - |
Photinus pyralis | AMP + diphosphate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000000157 | - |
MgATP2- | mutant enzyme K443A/K529A | Photinus pyralis | |
0.000000157 | - |
D-luciferin | mutant enzyme K443A/K529A | Photinus pyralis | |
0.0000311 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K443A/K529A | Photinus pyralis | |
0.0000629 | - |
MgATP2- | mutant enzyme K443A | Photinus pyralis | |
0.0000629 | - |
D-luciferin | mutant enzyme K443A | Photinus pyralis | |
0.000101 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K443A | Photinus pyralis | |
0.00025 | - |
MgATP2- | mutant enzyme K529A | Photinus pyralis | |
0.00025 | - |
D-luciferin | mutant enzyme K529A | Photinus pyralis | |
0.02 | - |
MgATP2- | mutant enzyme G446I | Photinus pyralis | |
0.02 | - |
D-luciferin | mutant enzyme G446I | Photinus pyralis | |
0.021 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme G446I | Photinus pyralis | |
0.0875 | - |
MgATP2- | mutant enzyme Q448A | Photinus pyralis | |
0.0875 | - |
D-luciferin | mutant enzyme Q448A | Photinus pyralis | |
0.095 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme Q448A | Photinus pyralis | |
0.167 | - |
MgATP2- | wild-type enzyme | Photinus pyralis | |
0.167 | - |
D-luciferin | wild-type enzyme | Photinus pyralis | |
0.175 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K529A | Photinus pyralis | |
0.23 | - |
D-luciferyl-O-adenosine monophosphate | wild-type enzyme | Photinus pyralis | |
0.24 | - |
MgATP2- | mutant enzyme K445Q | Photinus pyralis | |
0.24 | - |
D-luciferin | mutant enzyme K445Q | Photinus pyralis | |
0.32 | - |
D-luciferyl-O-adenosine monophosphate | mutant enzyme K445Q | Photinus pyralis |