Metallothionein-induced increase in mitochondrial inner membrane permeability.

Objective: To elucidate the effect of metallothionein on the permeability of the mitochondrial inner membrane.

Background: Metallothionein (MT) is a 6-7-kDa protein which is rapidly produced by stressed cells. MT is induced by cytokines and other factors thought to mediate the stress response. The organelle with the highest concentration of MT is the mitochondrion. Therefore we studied the effect of MT on mitochondrial function. We chose to study the effect of MT on mitochondrial inner membrane permeability because of the role of this function in numerous cellular processes.

Methods: Mitochondria were isolated from livers of Sprague-Dawley rats by differential centrifugation and suspended in sucrose-containing buffer. Changes in mitochondrial inner membrane permeability were monitored by following the change in absorbance at 540 nm. All experiments were of a paired design.

Results: We found that an increase in inner membrane permeability was induced by physiological metallothionein 1 (MT1) concentrations between 6 and 50 microM. There was no increase in the effect beyond 50 microM. The metals of MT1, zinc, and cadmium alone had no effect at physiological concentrations. The action of MT1 was inhibited by the aliphatic polyamine, spermine, as well as magnesium both at physiological concentrations. Spermine was effective whether added before or after MT1. Metallothionein 2 of different Zn2+ and Cd2+ compositions induced different kinetics of pore opening.

Conclusions: These experiments reveal the possibility that the permeability of the mitochondrial inner membrane is regulated by relative concentrations of MT, spermine, and magnesium. The metal composition of MT could also play a role in this regulation.