Intramolecular sensitization and structure of a Tb<sup>3+</sup>/2-hydroxyquinoline conjugate in the paraoxonase 1 active site.

Abstract

Paraoxonase 1 (PON1) is a Ca²⁺-dependent enzyme involved in oxidative stress processes and is widely studied for its protective roles in various diseases. Intermolecular sensitization of lanthanide ions was implemented by replacing Ca²⁺ ions from the recombinant PON1 (rePON1) catalytic site in the presence of 2-hydroxyquinoline (2HQ) as an external antenna. Although the replacement of Ca²⁺ ions with lanthanide ions indicates weaker binding affinity for the coordination of 2HQ in the protein milieu of the rePON1 active site, it results in the formation of a highly emissive supramolecular complex in the case of Tb³⁺ ions. The architecture of the ternary rePON1 : Tb³⁺ : 2HQ conjugate, which allows efficient terbium sensitization and its specific long-wavelength metal phosphorescence emission, was resolved by X-ray crystallography. These findings could establish a non-catalytic quantification strategy for PON1 and provide additional structural insights into lanthanide substitution in this Ca²⁺-dependent enzyme.