[Subunit interactions in luciferase from the firefly Luciola mingrelica. Their role in the manifestation of enzyme activity and during thermoinactivation].
It was shown that the dimers of the firefly luciferase possess the catalytic activity, whereas the monomers do not. The dissociation constant (Kd) for active dimers was determined at pH 7.0--8.4 within the temperature range of 15--35 degrees and at MgSO4 and Na2SO4 concentrations varying from 37 to 370 mM and 49 to 490 mM, respectively. Under variable conditions the Kd value changed only insignificantly and made up to 13 nm. The substitution of Na2SO4 for MgSO4 decreased Kd 2.5 times. The effective rate constant for the enzyme inactivation (kin) was increased more than 5-fold, when the luciferase concentration was decreased from 200 down to 3.5 nM in the presence of 37 mM MgSO4. When the concentration of the latter was increased up to 185 mM, the value of kin ceased to depend on the enzyme concentration. The decrease of kin was also observed at an increase in Na2SO4. An inactivation pattern for the enzyme in solution was determined both for the monomer and for the dimer of the enzyme. The equations allowing to calculate the inactivation constant for the monomer (Ki) and dimer (k2) at different pH values, temperatures and salt concentrations were obtained. The enzyme was found to be stabilized by salts more than 10-fold, the stabilizing effect being far more pronounced for the enzyme monomer than for the dimer. The dependence of the effective kin value on pH and temperature was primarily influenced by the dependence of the inactivation rate constant for the dimer.
D000818:Animals; D001517:Coleoptera / Q000201:enzymology*; D006358:Hot Temperature; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism*; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D009994:Osmolar Concentration; D012964:Sodium / Q000494:pharmacology
D046911:Macromolecular Substances; D012964:Sodium; D008156:Luciferases; D008274:Magnesium
Sub"edinichnye vzaimodeĭstviia v liutsiferaze svetliakov Luciola mingrelica. Ikh rol' v proiavlenii aktivnosti fermenta i protsesse termoinaktivatsii.