Characteristics of non-specific permeability and H+-ATPase inhibition induced in the plasma membrane of Nitella flexilis by excessive Cu2+

Effects of Cu2+ On a non-specific conductance and H+-ATPase activity in the plasma membrane of the freshwater alga Nitella flexilis L. Agardh was studied using a conventional microelectrode voltage-clamp technique. We show that a Cu2+-induced increase in the non-specific conductance is related to the formation of pores in the plasma membrane. Pore formation is the result of unidentified chemical reactions, since the Q(10) for the rate of increase of conductance over time was about 3. Various oxidants and antioxidants (10 mmol/l H2O2, 10 mmol/l ascorbate, 100 mug/ml superoxide dismutase, and 100 mug/ml catalase) did not alter Cu2+-induced changes in the plasma membrane conductance, suggesting that the effect of Cu2+ was unrelated to peroxidation of plasma-membrane lipids. In contrast, organic and inorganic Ca2+-channel antagonists (Zn2+, Cd2+, Fe2+, Ni2+) inhibited the Cu2+ induced non-specific conductance increase. This suggests that changes in Ca2+ influx underlie this effect of Cu2+. Decreasing the pH or the ionic strength of external solutions also inhibited the Cu2+-induced plasma-membrane conductance increase. Copper was also found to inhibit plasma-membrane H+-ATPase activity with half-maximal inhibition occurring at about 5-20 mu mol/l and full inhibition at about 100-300 mu mol/l. The Hill coefficient of Cu2+ inhibition of the H+-ATPase was close to two.