Electronic excitation transfer in the complex of lumazine protein with bacterial bioluminescence intermediates.
Fluorescence dynamics measurements have been made on the bioluminescence reaction intermediates using Photobacterium leiognathi, Vibrio fischeri, and Vibrio harveyi luciferases, both alone and in mixtures with Photobacterium phosphoreum lumazine protein. Each luciferase produces a "fluorescent transient" intermediate on reaction with the bioluminescence substrates, FMNH2, tetradecanal, and O2, and all have a fluorescence quantum yield about 0.3, with a predominant lifetime around 10 ns. The P. leiognathi luciferase fluorescent transient has a rotational correlation time of 79 ns at 2 degrees C, as expected for the rotational diffusion of a 77-kDa macromolecule. In the presence of lumazine protein however a faster correlation time of about 3 ns predominates. This rapid channel of anisotropy loss is attributed to energy transfer from the flavin intermediate bound on the luciferase to the lumazine ligand, reflects the presence of protein-protein complexation, and is greatest in the case of P. leiognathi, but not at all for V. fischeri. This fact is consistent with the strong influence of lumazine protein on the bioluminescence reaction of P. leiognathi, and not at all with V. fischeri. The rate of energy transfer is of order 10(9) s-1, much greater than the 10(8) s-1 fluorescence rate of the donor. Thus the bioluminescence excitation of lumazine protein could occur by a similar photophysical mechanism of interprotein energy transfer from a chemically excited fluorescent transient donor to the lumazine acceptor.
Lee
J
J
Department of Biochemistry, University of Georgia, Athens 30602.
Wang
Y Y
YY
Gibson
B G
BG
D001426:Bacterial Proteins / Q000378:metabolism*; D004735:Energy Transfer; D005454:Fluorescence Polarization; D008156:Luciferases; D008163:Luminescent Measurements; D010776:Photobacterium / Q000378:metabolism*; D011621:Pteridines / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014733:Vibrio / Q000378:metabolism*
D001426:Bacterial Proteins; D011621:Pteridines; C008297:lumazine; D008156:Luciferases