The inhibition of gadolinium ion (Gd3+) on the mitochondrial F1FO-ATPase is linked to the modulation of the mitochondrial permeability transition pore

Abstract

The mitochondrial permeability transition pore (PTP), which drives regulated cell death when Ca²⁺ concentration suddenly increases in mitochondria, was related to changes in the Ca²⁺-activated F₁F_O-ATPase. The effects of the gadolinium cation (Gd³⁺), widely used for diagnosis and therapy, and reported as PTP blocker, were evaluated on the F₁F_O-ATPase activated by Mg²⁺ or Ca²⁺ and on the PTP. Gd³⁺ more effectively inhibits the Ca²⁺-activated F₁F_O-ATPase than the Mg²⁺-activated F₁F_O-ATPase by a mixed-type inhibition on the former and by uncompetitive mechanism on the latter. Most likely Gd³⁺ binding to F₁, is favoured by Ca²⁺ insertion. The maximal inactivation rates (k_inact) of pseudo-first order inactivation are similar either when the F₁F_O-ATPase is activated by Ca²⁺ or by Mg²⁺. The half-maximal inactivator concentrations (K_I) are 2.35 ± 0.35 mM and 0.72 ± 0.11 mM, respectively. The potency of a mechanism-based inhibitor (k_inact/K_I) also highlights a higher inhibition efficiency of Gd³⁺ on the Ca²⁺-activated F₁F_O-ATPase (0.59 ± 0.09 mM^-1∙s^-1) than on the Mg²⁺-activated F₁F_O-ATPase (0.13 ± 0.02 mM^-1∙s^-1). Consistently, the PTP is desensitized in presence of Gd³⁺. The Gd³⁺ inhibition on both the mitochondrial Ca²⁺-activated F₁F_O-ATPase and the PTP strengthens the link between the PTP and the F₁F_O-ATPase when activated by Ca²⁺ and provides insights on the biological effects of Gd³⁺.

Keywords: Enzyme kinetics; F(1)F(O)-ATPase; Gadolinium ion; Metal cofactors; Mitochondria; Permeability transition pore.