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SENSITIZATION BY LUMAZINE PROTEINS OF THE BIOLUMINESCENCE EMISSION FROM THE REACTION OF BACTERIAL LUCIFERASES
Author(s) -
Lee John
Publication year - 1982
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1982.tb09490.x
Subject(s) - photobacterium phosphoreum , luciferases , bioluminescence , luciferase , vibrio harveyi , chemistry , photobacterium , stereochemistry , biochemistry , biology , vibrio , bacteria , organic chemistry , transfection , genetics , toxicity , gene
— Lumazine protein from Photobacterium phosphoreurn blue shifts the in vitro bioluminescence spectra in the reactions using each of the 4 main types of bacterial luciferases: P. phosphoreum, P. leiognathi, Vibrio harveyi and V. fischeri . For the reaction initiated with FMNH 2 and tetradecanal at 2°C, this “sensitizing” property of lumazine protein differs quantitatively between the luciferases. An interaction constant characterizing each type of luciferase may be derived from a reciprocal plot of the spectral shift against the lurnazine protein concentration. The weakest interaction constant is in the V. fischeri reaction, 180 μM. For the V. harveyi reaction the interaction is in the range 6–9 μ M , and for both Photobacterium reactions it is 2–3 μM. A concentration of only 0.6 μ M of lumazine protein is sufficient to cause an observable change in the Photobacterium bioluminescence spectra. For the V. harveyi case the interaction constant is near to the equilibrium K d for the luciferase‐lumazine protein complex, observed directly by Visser and Lee. Both constants are decreased markedly by increase in phosphate concentration so that it is concluded that, with V. harveyi luciferase, sensitization occurs within this protein‐protein complex. For P. phosphoreum luciferase, however, the equilibrium complex is too weak to correspond to the sensitizing interaction and it is concluded that the rate‐limiting process is a protein‐protein bimolecular collision. As judged from their molecular weight around 20000, spectral properties, and ability to blue shift the bioluminescence spectra, lumazine proteins are identified in a second strain of P. phosphoreum and in P. leiognathi .