Occurrence of P-flavin binding protein in Vibrio fischeri and properties of the protein.
In previous studies involving Photobacterium species we proposed that (i) P-flavin is the product of luciferase, (ii) the physiological function of the lux operon is not to produce light but to produce FP(390) (luxF protein), including its prosthetic group, P-flavin, and (iii) FP(390) reactivates oxidatively inactivated cobalamin-dependent methionine synthase similar to flavodoxin but at relatively high ionic strength. It seems difficult to extend this idea to all luminous bacteria because the luxF gene is not present in the lux operon in Vibrio or Xenorhabdus. But we predicted that a luciferase fragment which binds P-flavin should function like FP(390) in these species. In this study, we isolated P-flavin binding protein from Vibrio fischeri ATCC 7744. The obtained protein was a modified luciferase as expected, in which the beta-subunit was intact but about 25 amino acid residues at the C-terminus of the alpha-subunit were deleted and the prosthetic group was the fully reduced P-flavin. These results strongly support that the physiological function of the lux operon is as described above even in luminous bacteria other than Photobacterium species. We propose that chromophore B reported by Tu and Hastings [Tu, S.-C. and Hastings, J.W. (1975) Biochemistry 14, 1975-1980] is the reduced P-flavin.
Kasai
S
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Department of Bioapplied Chemistry, Faculty of Engineering, Osaka City University, Sumiyoshi-ku, Osaka, 558-8585, Japan. kasai@bioa.eng.osaka-cu.ac.jp
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D002352:Carrier Proteins / Q000378:metabolism; D002847:Chromatography, Agarose; D002855:Chromatography, Thin Layer; D005415:Flavins / Q000737:chemistry* / Q000378:metabolism*; D005418:Flavodoxin / Q000378:metabolism; D008156:Luciferases / Q000737:chemistry / Q000302:isolation & purification; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D014733:Vibrio / Q000737:chemistry*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D005415:Flavins; D005418:Flavodoxin; D010446:Peptide Fragments; D008156:Luciferases