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Related Experiment Videos

Borrowed proteins in bacterial bioluminescence.

D J O'Kane1, B Woodward, J Lee

  • 1Department of Biochemistry, University of Georgia, Athens 30602.

Proceedings of the National Academy of Sciences of the United States of America
|February 15, 1991
PubMed
Summary

Researchers identified the lumazine protein gene in Photobacterium phosphoreum. Sequence analysis revealed similarities to other proteins, suggesting a conserved lumazine binding site structure.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Photobacterium phosphoreum is a marine bacterium known for bioluminescence.
  • Lumazine protein plays a role in light production within certain bacteria.
  • Understanding lumazine protein function requires gene identification and sequence analysis.

Purpose of the Study:

  • To isolate and characterize the lumazine protein gene from Photobacterium phosphoreum.
  • To determine the amino acid sequence of the lumazine protein.
  • To investigate potential functional and structural similarities with related proteins.

Main Methods:

  • Screening a Photobacterium phosphoreum DNA library using degenerate oligonucleotide probes.
  • Gene localization to a specific DNA fragment (3.4-kilobase BamHI/EcoRI).

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  • Sequence analysis of the open reading frame and comparison with known protein sequences.

    • Main Results:

    • The lumazine protein gene was successfully identified and localized.
    • A 189-residue protein sequence was predicted, consistent with partial experimental data.
    • Significant sequence similarity was found between lumazine protein, Vibrio fischeri yellow fluorescence protein, and riboflavin synthetase alpha subunit.
    • A conserved sequence in lumazine protein aligns with proposed lumazine binding sites.

    Conclusions:

    • The study successfully cloned and sequenced the lumazine protein gene from Photobacterium phosphoreum.
    • Structural and functional homology suggests a conserved lumazine binding mechanism across related proteins.
    • A proposed beta-sheet structure for the lumazine binding site is supported by computational predictions and experimental data.