Display Settings:

Format

Send to:

Choose Destination
    Proc Natl Acad Sci U S A. 1973 Dec;70(12 Pt 1-2):3468-72.

    Spectral properties of an oxygenated luciferase-flavin intermediate isolated by low-temperature chromatography.

    Source

    Ecole des Hautes Etudes, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, Paris 5 France.

    Abstract

    Bacterial luciferase catalyzes the oxidation of reduced flavin mononucleotide by molecular oxygen; long-chain aldehyde is required for light emission. At 20 degrees the bioluminescence has a lifetime of tens of seconds, while excess reduced flavin is removed by way of nonenzymatic autoxidation in less than a second. This observation indicates the existence of a long-lived enzyme intermediate, which has been postulated to be a peroxide of the enzyme-bound reduced flavin. This intermediate was isolated and studied at low temperature (-20 degrees ), where it has a lifetime measured in days. It has an absorption with a single band peaking at 372 nm, and fluorescence emission centered at about 485 nm, which might be expected for the postulated flavin peroxide. Upon conversion to product, flavin mononucleotide-like absorption and fluorescence appear, supporting the postulate that flavin turns over in the reaction. Upon injection into buffer at 20 degrees with added aldehyde, bioluminescence occurs. Based on a stoichiometry of one flavin per luciferase molecule, the specific activity of the intermediate is equal to that of pure luciferase.

    PMID:
    16592121
    [PubMed]
    PMCID:
    PMC427261
    Free PMC Article

      Supplemental Content

      Icon for PubMed Central

      Save items

      loading

      Recent activity

      Your browsing activity is empty.

      Activity recording is turned off.

      Turn recording back on

      See more...
      Write to the Help Desk