Literature summary for 1.14.14.3 extracted from

Campbell, Z.T.; Weichsel, A.; Montfort, W.R.; Baldwin, T.O.
Crystal structure of the bacterial luciferase/flavin complex provides insight into the function of the beta subunit (2009), Biochemistry, 48, 6085-6094.

Search for more literature for 1.14.14.3

Cloned(Commentary)

Cloned (Comment)	Organism
amplified from the pJHD500 plasmid and ligated into a pET21b vector, expressed from pZCH2 in an Escherichia coli BL21 (lambdaDE3) cell line after growth to an OD600 of 0.5	Vibrio harveyi

Crystallization (Commentary)

Crystallization (Comment)	Organism
bacterial luciferase/FMN complex, by the hanging drop method, at 2.3 A resolution. Crystals of recombinant luciferase are grown at room temperature prior to soaking with millimolar concentrations of FMN. Belongs to space group P212121. The isoalloxazine ring is coordinated by an unusual cis-Ala-Ala peptide bond. The reactive sulfhydryl group of Cys106 projects toward position C-4a, the site of flavin oxygenation. Mobile loop that is crystallographically disordered, appears to be a boundary between solvent and the active center. Within this portion of the protein, there is a single contact between Phe272 of the R subunit and Tyr151 of the beta subunit	Vibrio harveyi

Crystallization (Comment)

Organism

bacterial luciferase/FMN complex, by the hanging drop method, at 2.3 A resolution. Crystals of recombinant luciferase are grown at room temperature prior to soaking with millimolar concentrations of FMN. Belongs to space group P212121. The isoalloxazine ring is coordinated by an unusual cis-Ala-Ala peptide bond. The reactive sulfhydryl group of Cys106 projects toward position C-4a, the site of flavin oxygenation. Mobile loop that is crystallographically disordered, appears to be a boundary between solvent and the active center. Within this portion of the protein, there is a single contact between Phe272 of the R subunit and Tyr151 of the beta subunit

Vibrio harveyi

Protein Variants

Protein Variants	Comment	Organism
Y151A	binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi
Y151D	binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi
Y151K	binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi
Y151R	binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi
Y151T	binds FMNH2 more weakly in comparison to the wild-type, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi
Y151W	least active mutant, binds reduced flavin with wild-type affinity, substitution at position 151 on the beta subunit causes reductions in activity and total quantum yield	Vibrio harveyi

Organism

Organism	UniProt	Comment	Textmining
Vibrio harveyi	P07740	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
37	-	wild-type enzyme has a 90% reduction in activity after 92 min. Comparable loss for the Y151W mutant after only 11 min	Vibrio harveyi

Cofactor

Cofactor	Comment	Organism	Structure
FMN	-	Vibrio harveyi