Abstract

The characteristics of (6,7-dimethyl-8-ribityl) lumazine proteins isolated from the marine bioluminescent bacteria Photobacterium phosphoreum and P. leiognathi, will be determined. The primary structure of each of these proteins will be determined using classical methods. Fast preparative HPLC methods will be developed to purify these proteins together with the luciferases from Photobacterium and bioluminescent Vibrios. Spectroscopic techniques using a Picosecond Photon Counting Spectrometer will be employed to measure the parameters of the association of lumazine protein and its ligand, and the lumazine protein-luciferase associations. Other ligands will be attached to lumazine protein instead of the lumazine derivative and these artificial proteins will be examined for their fluorescence and bioluminescence properties. Energy transfer techniques will be used to describe the topological features of the protein-protein complexes. A search will be made for lumazine proteins in bioluminescent Vibrios as well as in other non-bioluminescent procaryotes. 6,7-dimethyl-8-ribityllumazine is an intermediary metabolite in Vitamin B2 biosynthesis and study of its binding to proteins may lead to the finding of new types of broad spectrum antibiotics.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM028139-07
Application #
3275393
Study Section
Biophysics and Biophysical Chemistry A Study Section (BBCA)
Project Start
1979-12-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Georgia
Department
Type
Schools of Arts and Sciences
DUNS #
City
Athens
State
GA
Country
United States
Zip Code
30602

  Publications

Petushkov, V N; Ketelaars, M; Gibson, B G et al. (1996) Interaction of Photobacterium leiognathi and Vibrio fischeri Y1 luciferases with fluorescent (antenna) proteins: bioluminescence effects of the aliphatic additive. Biochemistry 35:12086-93
Petushkov, V N; Gibson, B G; Lee, J (1996) Direct measurement of excitation transfer in the protein complex of bacterial luciferase hydroxyflavin and the associated yellow fluorescence proteins from Vibrio fischeri Y1. Biochemistry 35:8413-8
Petushkov, V N; Gibson, B G; Lee, J (1995) Properties of recombinant fluorescent proteins from Photobacterium leiognathi and their interaction with luciferase intermediates. Biochemistry 34:3300-9
Illarionov, B; Illarionova, V; Lee, J et al. (1994) Expression and properties of the recombinant lumazine (riboflavin) protein from Photobacterium leiognathi. Biochim Biophys Acta 1201:251-8
Moore, S A; James, M N; O'Kane, D J et al. (1993) Crystal structure of a flavoprotein related to the subunits of bacterial luciferase. EMBO J 12:1767-74
Lee, J (1993) Lumazine protein and the excitation mechanism in bacterial bioluminescence. Biophys Chem 48:149-58
Lee, J; Gibson, B G; O'Kane, D J et al. (1992) Fluorescence study of the ligand stereospecificity for binding to lumazine protein. Eur J Biochem 210:711-9
O'Kane, D J; Prasher, D C (1992) Evolutionary origins of bacterial bioluminescence. Mol Microbiol 6:443-9
Lee, J; Wang, Y Y; Gibson, B G (1991) Electronic excitation transfer in the complex of lumazine protein with bacterial bioluminescence intermediates. Biochemistry 30:6825-35
O'Kane, D J; Woodward, B; Lee, J et al. (1991) Borrowed proteins in bacterial bioluminescence. Proc Natl Acad Sci U S A 88:1100-4

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