Bioluminescence emission of bacterial luciferase with 1-deaza-FMN. Evidence for the noninvolvement of N(1)-protonated flavin species as emitters.
The reaction of reduced 1-d-FMN with oxygen and decanal results in bioluminescence with kinetic and spectral properties similar to those of the reaction with FMNH2, even though the spectral (absorbance, fluorescence) and chemical properties of the oxidized forms differ greatly. This emission, which is about 10-15% as efficient as with FMNH2, is postulated to involve the intermediacy of the corresponding 4a-hydroperoxide, the fluorescence of which occurred transiently. The N(1) protonated species had been proposed as the emitter in the reaction with FMNH2, but the 1-deaza analog cannot be protonated at the corresponding position, thus excluding this possibility.
Kurfürst
M
M
Fakultät für Biologie der Universität, Konstanz, Federal Republic of Germany.
Macheroux
P
P
Ghisla
S
S
Hastings
J W
JW
D005486:Flavin Mononucleotide / Q000031:analogs & derivatives* / Q000378:metabolism; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D014733:Vibrio / Q000201:enzymology*
C059658:1-deaza-FMN; D005486:Flavin Mononucleotide; D008156:Luciferases