Isolation of the in vivo emitter in bacterial bioluminescence
- 1 February 1978
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 75 (2) , 833-837
- https://doi.org/10.1073/pnas.75.2.833
Abstract
A blue fluorescence protein has been isolated and purified from extracts of the luminous bacterium Photobacterium phosphoreum . It is a single polypeptide of molecular weight 22,000 with absorption maxima at 274 and 418 nm. It is efficiently fluorescent (ϕ F 0.45), with a fully corrected spectral maximum (476 nm) and distribution identical to the in vivo bioluminescence from this same type of bacterium. At low concentration this fluorescence shifts towards the red and becomes identical to the in vitro bioluminescence emission. This spectral shift apparently results from a change in the protein pulled by dissociation of the chromophore ( K d [unk] 10 -7 M). If the blue fluorescence protein is included in the in vitro bioluminescence reaction with reduced FMN, oxygen, aldehyde, and luciferase ( P. phosphoreum ), the bioluminescence spectrum is shifted towards the blue from its maximum at 490 nm to one at 476 nm, where it is again identical in all respects to the in vivo bioluminescence spectrum. This is accompanied by an increase in the initial light intensity by an order of magnitude at saturating levels of blue fluorescence protein, and the specific light yield of the luciferase is increased 4-fold. It is suggested that the blue fluorescence protein acts as a sensitizer of the bacterial bioluminescence reaction.Keywords
This publication has 26 references indexed in Scilit:
- Photoexcited bacterial luminescence. Spectral properties and mechanistic implication of a reduced flavine-like prosthetic group associated with photoexcitable luciferaseBiochemistry, 1975
- Bacterial bioluminescence. Equilibrium association measurements, quantum yields, reaction kinetics, and overall reaction schemeBiochemistry, 1975
- Separation of the apoprotein and reconstitution of the holoprotein from the long-lived intermediate in bacterial bioluminescenceBiochemical and Biophysical Research Communications, 1974
- Bacterial bioluminescence - identification of fatty acid as product, its quantum yield and a suggested mechanismBiochemical and Biophysical Research Communications, 1973
- Bacterial bioluminescence. Quantum yields and stoichiometry of the reactants reduced flavin mononucleotide, dodecanal, and oxygen, and of a product hydrogen peroxideBiochemistry, 1972
- BioluminescenceAnnual Review of Biochemistry, 1964
- Investigations on the identity of the light-emitting molecule in Photobacterium phosphoreumBiochimica et Biophysica Acta, 1963
- Studies on the bioluminescence of Renilla reniformis III. Some biochemical comparisons of the system to other Renilla species and determination of the spectral energy distributionsBiochimica et Biophysica Acta, 1962
- Evidence for the presence of an unknown factor, active in the light reaction, in preparations of Photobacterium phosphoreumBiochimica et Biophysica Acta, 1962
- Emission spectra of luminous bacteriaBiochimica et Biophysica Acta, 1950