Role of metal ions in triazine dye affinity chromatography: The metal mediated interaction of triazine dyes with firefly luciferase

Low concentrations of metals of the first row transition series, Zn²⁺, Co²⁺, Mn²⁺, Ni²⁺, Cu²⁺ and Fe²⁺, and to a lesser extent the group IIa ions, particularly Mg²⁺, influenced the interaction of firefly luciferase Photinus luciferin:oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolysing), EC 1.13.12.7] with a number of triazine dyes. For example Cu²⁺ promoted the binding of luciferase to Cibacron Brilliant Blue (BR-II) and Cibacron Blue F3GA a dichloro and monochloro triazine dye, respectively. On the other hand Zn²⁺ prevented dye inactivation and even enhanced the enzyme activity. Specificity was observed in the interference of different metals interacting with different dye-protein. This is made use of in triazine dye affinity chromatography.