coelenterazine and sodium-thiosulfate

The light emission of obelin may be initiated by Mn2+ under alkaline conditions. The luminescence takes place in a pH range from 7 to 12 with a sharp optimum at 11.75. The first-order rate constant for Mn(2+)-activated luminescence decay is more than 9 s-1, while that for Ca(2+)-activated luminescence decay is only 6.9 s-1. The Mn2+ concentration-effect curve for obelin determined with simple dilutions of manganese salt is a sigmoid curve. The slope of the curve is moderately dependent on the pH and was not more than 1 within the pH range tested. The maximal light emission, which is initiated by 3.6 x 10(-5) M Mn2+ at pH 11.75 was about 10% of the maximal Ca(2+)-activated luminescence. Mg2+ ions inhibit the Mn(2+)-activated luminescence of obelin. The addition of OH. and O2- scavengers did not influence the Mn(2+)-activated luminescence, but when singlet oxygen quenchers were added, the Mn(2+)-dependent light emission was inhibited. This suggests that the 1O2 might be formed and itself be responsible for chromophore oxidation attended with light emission. NEM and Na2S2O4 inhibit the Mn(2+)-initiated light emission of obelin completely, showing that endogenous hydroperoxide and SH-group(s) of the photoprotein are essential for both Ca(2+)-activated and Mn(2+)-activated light emission of obelin.

Topics: Aequorin; Animals; Cnidaria; Dose-Response Relationship, Drug; Free Radical Scavengers; Free Radicals; Hydrogen-Ion Concentration; Imidazoles; Kinetics; Light; Luminescent Measurements; Luminescent Proteins; Manganese; Models, Chemical; Pyrazines; Recombinant Proteins; Sulfhydryl Reagents; Thiosulfates