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Literature summary for 1.14.14.3 extracted from

  • Campbell, Z.T.; Baldwin, T.O.
    Two lysine residues in the bacterial luciferase mobile loop stabilize reaction intermediates (2009), J. Biol. Chem., 284, 32827-32834.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.14.3

Cloned(Commentary)

Cloned (Comment) Organism
ligated into pET21-b vector, expressed from pZCH2 in Escherichia coli BL21 (lambdaDE3) cell line Vibrio harveyi

Protein Variants

Protein Variants Comment Organism
D262A 90% reduced activity with octanal, 36% reduced activity with decanal, activity with dodecanal as the wild-type Vibrio harveyi
D265A activity with octanal as the wild-type, 81% reduced activity with decanal, complete loss of dodecanal activity Vibrio harveyi
D271A complete loss of octanal and decanal activity, 18% reduced activity with dodecanal Vibrio harveyi
H285A 26% reduced activity with octanal, 74% reduced activity with decanal, complete loss of dodecanal activity Vibrio harveyi
K274A 89% reduced activity with octanal, 21% reduced activity with decanal, 81% reduced activity with dodecanal Vibrio harveyi
K283A complete loss of octanal and decanal activity, 96% reduced activity with dodecanal, does not significantly impede binding of decanal, results in destabilization of intermediate II, results in a loss in quantum yield comparable with that of the loop deletion mutant, binds reduced flavin more weakly Vibrio harveyi
K286A 92% reduced activity with octanal, complete loss of decanal activity, 87% reduced activity with dodecanal, does not significantly impede binding of decanal, increase in exposure of reaction intermediates to a dynamic quencher, results in a loss in quantum yield comparable with that of the loop deletion mutant, binds reduced flavin more weakly Vibrio harveyi
R291A 77% reduced activity with octanal, 58% reduced activity with decanal, 71% reduced activity with dodecanal Vibrio harveyi
W277A 11% reduced activity with octanal, 50% reduced activity with decanal and dodecanal Vibrio harveyi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.008
-
 decanal wild-type Vibrio harveyi
0.01
-
 decanal mutant K286A Vibrio harveyi
0.0105
-
 decanal mutant K283A Vibrio harveyi

Organism

Organism UniProt Comment Textmining
Vibrio harveyi
-
-
-

Purification (Commentary)

Purification (Comment) Organism
on nickel affinity column, to more than 90% purity Vibrio harveyi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
decanal + FMNH + O2
-
 Vibrio harveyi decanoic acid + FMN + H2O + light
-
 ?
dodecanal + FMNH + O2
-
 Vibrio harveyi dodecanoic acid + FMN + H2O + light
-
 ?
octanal + FMNH + O2
-
 Vibrio harveyi octanoic acid + FMN + H2O + light
-
 ?

Synonyms

Synonyms Comment Organism
bacterial luciferase
-
 Vibrio harveyi

Cofactor

Cofactor Comment Organism Structure
FMN
-
 Vibrio harveyi