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Literature summary extracted from

  • Branchini, B.R.; Southworth, T.L.; Murtiashaw, M.H.; Boije, H.; Fleet, S.E.
    A mutagenesis study of the putative luciferin binding site residues of firefly luciferase (2003), Biochemistry, 42, 10429-10436.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.13.12.7 mutant enzymes,R218A, H245A, G246A, F247A, F247L, F247Y, F250G, F250S, T251A, G315A, G316A, G341A, L342A, T343A, S347A, A348V, I351A and K529A Photinus pyralis

Protein Variants

EC Number Protein Variants Comment Organism
1.13.12.7 A348V Km-value for D-luciferin is 8.9fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 2fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 6.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 F247A Km-value for D-luciferin is 15.3fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 17.9fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 21.9fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 F247L Km-value for D-luciferin is 8.3fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is nearly identical to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.4fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 F247Y Km-value for D-luciferin is 1.2fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 128fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 2fold lower compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 F250G Km-value for D-luciferin is neraly identical to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 9.6fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is nearly identical to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 F250S Km-value for D-luciferin is 1.5fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 3.5fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.2fold lower compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 G246A Km-value for D-luciferin is 3.6fold lower compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 2.1fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 5.5fold lower compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 G315A Km-value for D-luciferin is 13.3fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 625fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 7.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 G316A Km-value for D-luciferin is 1.2fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 2.6fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.5fold lower compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 G341A Km-value for D-luciferin is 4fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 625fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.3fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 H245A Km-value for D-luciferin is identical to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 3fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 I351A Km-value for D-luciferin is 5.7fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 1.5fold higher compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 1.8fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 K529A Km-value for D-luciferin is 15.3fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 1250fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 7.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 L342A Km-value for D-luciferin is 8.7fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 1.2fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 7.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 R218A Km-value for D-luciferin is 20fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 31.3fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 38.8fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 S347A Km-value for D-luciferin is 11.3fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 4.2fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 2.2fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 T251A Km-value for D-luciferin is 20.7fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 2.9fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 12.5fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis
1.13.12.7 T343A Km-value for D-luciferin is 6.6fold higher compared to the Km-value of the wild-type enzyme, the turnover-number for D-luciferin is 125fold lower compared to the turnover-number of the wild-type enzyme, the Km-value for MgATP2- is 5.4fold higher compared to the Km-value of the wild-type enzyme Photinus pyralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.13.12.7 0.0042
-
 D-luciferin mutant enzyme G246A Photinus pyralis
1.13.12.7 0.015
-
 D-luciferin wild-type enzyme Photinus pyralis
1.13.12.7 0.015
-
 D-luciferin mutant enzyme H245A Photinus pyralis
1.13.12.7 0.0157
-
 D-luciferin mutant enzyme F250G Photinus pyralis
1.13.12.7 0.018
-
 D-luciferin mutant enzyme F247Y Photinus pyralis
1.13.12.7 0.018
-
 D-luciferin mutant enzyme G316A Photinus pyralis
1.13.12.7 0.022
-
 D-luciferin mutant enzyme F250S Photinus pyralis
1.13.12.7 0.029
-
 MgATP2- mutant enzyme G246A Photinus pyralis
1.13.12.7 0.06
-
 D-luciferin mutant enzyme G341A Photinus pyralis
1.13.12.7 0.08
-
 MgATP2- mutant enzyme F247Y Photinus pyralis
1.13.12.7 0.085
-
 D-luciferin mutant enzyme I351A Photinus pyralis
1.13.12.7 0.099
-
 D-luciferin mutant enzyme T343A Photinus pyralis
1.13.12.7 0.107
-
 MgATP2- mutant enzyme G316A Photinus pyralis
1.13.12.7 0.124
-
 D-luciferin mutant enzyme F247L Photinus pyralis
1.13.12.7 0.13
-
 D-luciferin mutant enzyme L342A Photinus pyralis
1.13.12.7 0.133
-
 D-luciferin mutant enzyme A348V Photinus pyralis
1.13.12.7 0.133
-
 MgATP2- mutant enzyme F250S Photinus pyralis
1.13.12.7 0.16
-
 MgATP2- wild-type enzyme Photinus pyralis
1.13.12.7 0.166
-
 MgATP2- mutant enzyme F250G Photinus pyralis
1.13.12.7 0.17
-
 D-luciferin mutant enzyme S347A Photinus pyralis
1.13.12.7 0.2
-
 D-luciferin mutant enzyme G315A Photinus pyralis
1.13.12.7 0.2
-
 MgATP2- mutant enzyme G341A Photinus pyralis
1.13.12.7 0.226
-
 MgATP2- mutant enzyme F247L Photinus pyralis
1.13.12.7 0.23
-
 D-luciferin mutant enzyme F247A Photinus pyralis
1.13.12.7 0.23
-
 D-luciferin mutant enzyme K529A Photinus pyralis
1.13.12.7 0.24
-
 MgATP2- mutant enzyme H245A Photinus pyralis
1.13.12.7 0.285
-
 MgATP2- mutant enzyme I351A Photinus pyralis
1.13.12.7 0.301
-
 D-luciferin mutant enzyme R218A Photinus pyralis
1.13.12.7 0.311
-
 D-luciferin mutant enzyme T251A Photinus pyralis
1.13.12.7 0.344
-
 MgATP2- mutant enzyme S347A Photinus pyralis
1.13.12.7 0.857
-
 MgATP2- mutant enzyme T343A Photinus pyralis
1.13.12.7 1.038
-
 MgATP2- mutant enzyme A348V Photinus pyralis
1.13.12.7 1.2
-
 MgATP2- mutant enzyme G315A Photinus pyralis
1.13.12.7 1.2
-
 MgATP2- mutant enzyme K529A Photinus pyralis
1.13.12.7 1.2
-
 MgATP2- mutant enzyme L342A Photinus pyralis
1.13.12.7 2
-
 MgATP2- mutant enzyme T251A Photinus pyralis
1.13.12.7 3.5
-
 MgATP2- mutant enzyme F247A Photinus pyralis
1.13.12.7 6.2
-
 MgATP2- mutant enzyme R218A Photinus pyralis

Organism

EC Number Organism UniProt Comment Textmining
1.13.12.7 Photinus pyralis
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.13.12.7 D-luciferin + O2 + ATP
-
 Photinus pyralis oxidized luciferin + CO2 + H2O + AMP + diphosphate + hv
-
 ?
1.13.12.7 luciferin + MgATP2-
-
 Photinus pyralis AMP + diphosphate
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.13.12.7 0.0001
-
 D-luciferin mutant enzyme K529A Photinus pyralis
1.13.12.7 0.0002
-
 D-luciferin mutant enzyme G315A Photinus pyralis
1.13.12.7 0.0002
-
 D-luciferin mutant enzyme G341A Photinus pyralis
1.13.12.7 0.001
-
 D-luciferin mutant enzyme T343A Photinus pyralis
1.13.12.7 0.004
-
 D-luciferin mutant enzyme R218A Photinus pyralis
1.13.12.7 0.007
-
 D-luciferin mutant enzyme F247A Photinus pyralis
1.13.12.7 0.013
-
 D-luciferin mutant enzyme F250G Photinus pyralis
1.13.12.7 0.03
-
 D-luciferin mutant enzyme S347A Photinus pyralis
1.13.12.7 0.036
-
 D-luciferin mutant enzyme F250S Photinus pyralis
1.13.12.7 0.041
-
 D-luciferin mutant enzyme H245A Photinus pyralis
1.13.12.7 0.043
-
 D-luciferin mutant enzyme T251A Photinus pyralis
1.13.12.7 0.049
-
 D-luciferin mutant enzyme G316A Photinus pyralis
1.13.12.7 0.06
-
 D-luciferin mutant enzyme G246A Photinus pyralis
1.13.12.7 0.064
-
 D-luciferin mutant enzyme A348V Photinus pyralis
1.13.12.7 0.098
-
 D-luciferin mutant enzyme F247Y Photinus pyralis
1.13.12.7 0.106
-
 D-luciferin mutant enzyme L342A Photinus pyralis
1.13.12.7 0.116
-
 D-luciferin mutant enzyme F247L Photinus pyralis
1.13.12.7 0.125
-
 D-luciferin wild-type enzyme Photinus pyralis
1.13.12.7 0.19
-
 D-luciferin mutant enzyme I351A Photinus pyralis