Time-resolved fluorescence spectroscopy of lumazine protein from Photobacterium phosphoreum using synchrotron radiation

  title={Time-resolved fluorescence spectroscopy of lumazine protein from Photobacterium phosphoreum using synchrotron radiation},
  author={Antonie J.W.G. Visser and Arie van Hoek and Dennis J. O'Kane and J. Lee},
  journal={European Biophysics Journal},
  • A. VisserA. Hoek J. Lee
  • Published 1 June 1989
  • Chemistry, Physics, Biology
  • European Biophysics Journal
Time-resolved fluorescence on lumazine protein from Photobacterium phosphoreum was performed with synchrotron radiation as a source of continuously tunable excitation. The experiments yielded structural and dynamic details from which two aspects became apparent. From fluorescence anisotropy decay monitoring of lumazine fluorescence with different excitation wavelengths, the average correlation times were shown to change, which must indicate the presence of anisotropic motion of the protein. A… 
7 Citations

Conformational studies of a hyperthermostable enzyme

The structural features of the hyperthermophilic endo‐β‐1,3‐glucanase from Pyrococcus furiosus were studied using circular dichroism, steady‐state and time‐resolved fluorescence spectroscopy and anisotropy to show spectral differences arose from changes in the local environment around specific tryptophan residues in the native, partially folded, partially unfolded and completely unfolded state.

Shape and lipid-binding site of the nonspecific lipid-transfer protein (sterol carrier protein 2): a steady-state and time-resolved fluorescence study.

The nonspecific lipid-transfer protein (nsL-TP) from bovine liver was studied with time-resolved and steady-state fluorescence techniques and it is concluded that nsL- TP is highly asymmetric.

Dissociation of lactate dehydrogenase in aqueous and reversed micellar solutions. A time-resolved polarized fluorescence study.

Dissociation behavior of lactate dehydrogenase from hog muscle, both in aqueous solution and reversed micelles of sodium bis(2-ethylhexyl sulfosuccinate) in octane was studied using time-resolved

Evolutionary origins of bacterial bioluminescence

Models describing the evolution of these paralogous proteins are suggested, as well as a postulate for the identity of the gene encoding a protobioluminescent luciferase are suggested.

Fluorescence anisotropy decay study of self-association of bacterial luciferase intermediates

The fluorescence dynamics parameters of the fluorescent transient flavin-luciferase species from the typesVibrio fischeri andPhotobacterium leiognathi are presented and it is proposed that the self-association competes with the lumazine protein interaction in the bioluminescence reaction.

Time‐Resolved Fluorescence Study of the Dissociation of FMN from the Yellow Fluorescence Protein from Vibrio fischeri

Time-resolved fluorescence spectroscopy of the flavin mononucleotide (FMN) prosthetic group of the yellow fluorescence protein (YFP) from Vibrio @hen has provided quantitative, thermodynamic



Purification of lumazine proteins from Photobacterium leiognathi and Photobacterium phosphoreum: bioluminescence properties.

Bright strains of the marine bioluminescent bacterium Photobacterium leiognathi produce a "lumazine protein" in amounts comparable to that previously found in Photobacteria phosphoreum, but in dimmer strains the amounts of lumazine protein in extracts are less, and also there is an accompanying shift of the biolumscence spectral maximum to longer wavelength.

Spectral properties and function of two lumazine proteins from Photobacterium.

The spectral properties are compared for two 6,7-dimethyl-8-ribityllumazine proteins from marine bioluminescent bacteria, one from a psychrophile, Photobacterium phosphoreum, and the other from a

Determination of rotational correlation times from deconvoluted fluorescence anisotropy decay curves. Demonstration with 6,7-dimethyl-8-ribityllumazine and lumazine protein from Photobacterium leiognathi as fluorescent indicators.

The experimental and analytical protocols required for obtaining rotational correlation times of biological macromolecules from fluorescence anisotropy decay measurements are described and the unbound 6,7-dimethyl-8-ribityllumazine protein was used as a model compound for determining correlation times in the picosecond time range.

Physical characterization of lumazine proteins from Photobacterium.

The physicochemical properties of Photobacterium lumazine proteins have been investigated, and there is insufficient area on the exterior surface to accommodate hydration when the lumzine proteins are considered as smooth-surfaced ellipsoids.

Spectroscopic investigations of the single tryptophan residue and of riboflavin and 7-oxolumazine bound to lumazine apoprotein from Photobacterium leiognathi.

A new method is designed for evaluation of the rate constant of energy transfer by measuring the (picosecond) rise time of the acceptor fluorescence, which indicates no change in secondary structure on binding to the apoprotein.

Dynamic fluorescence properties of bacterial luciferase intermediates.

The results favor a proposal that, in these intermediate states, the luciferase undergoes a conformational change in which its axial ratio increases by 50%.

Chemical characterization of lumazine protein from Photobacterium leiognathi: comparison with lumazine protein from Photobacterium phosphoreum.

The UV and visible absorption spectra of both lumazine proteins denatured in 6 M guanidine hydrochloride can be accurately modeled by using the number of equivalents of the lumazine derivative and blocked aromatic amino acid model compounds determined by chemical and spectrophotometric analyses for Trp, Tyr, and Phe.

Application of a reference convolution method to tryptophan fluorescence in proteins. A refined description of rotational dynamics.

Fluorescence lifetime determinations for p-terphenyl, p-bis[2-(5-phenyloxazolyl)]benzene and N-acetyltryptophanamide show that with this method more reliable fits of the decays can be made than with the scatterer method, which is most frequently used.

Fluorescence decay studies of anisotropic rotations of small molecules

The fluorescence decay and the decay of the emission anisotropy of perylene and 9‐aminoacridine in glycerol have been investigated at several excitation wavelengths over the temperature range 10 to

Fluorescence depolarization studies on the flexibility of myosin rod.

It is concluded that, at pH 8, the hinge region of the myosin rods has considerable resistance to bending, more like a spring than a free hinge.