Cadmium interactions with ATPase activity in the euryhaline alga Dunaliella bioculata

Abstract

To further study the toxicity of cadmium in the euryhaline alga, Dunaliella bioculata, ATPase activity and Cd²⁺ interactions were investigated in this species.

Ultracytochemical studies showed the presence of ATPase reaction after incubation with Ca²⁺ and Mg²⁺, on different cell structures, the cytoplasm, the nucleoplasm, the axoneme and the membrane of the flagellae. In the cytoplasm, the localization of the lead precipates suggests that they are associated with the endoplasmic reticulum.

The in vitro measurement of enzyme activity in crude cell extracts obtained by a partial solubilization of deflagellated algae with Triton X100, revealed a high Mg²⁺ dependent pyrophosphatase activity, a weak Mg²⁺-ATPase and a Ca²⁺-ATPase (Km = 0.12 mM) which was little sensitive to vanadate. In these extracts, a Ca²⁺ dependent ATPase was detected at the level of a double band by a non-denaturing electrophoresis. The same activity was found in the supernatant of sonicated cells in the absence of detergent, which suggests that this ATPase could be a cytosolic enzyme.

In plasma membrane fractions, vanadate-sensitive ATPase activity was measured. This reaction was activated either by Mg²⁺ at relatively low concentrations (Km = 150µm) or by Ca² +, but required unusually high concentrations of this ion, 50–100 mM.

The inhibitory effects of Cd²⁺ on Ca²⁺ ATPase activity in cell extracts were compared with those of other cations. The range of toxicity was: Zn²⁺ > Cd²⁺ > Cu²⁺ > La³⁺ > Co²⁺. For Cd²⁺, the IC₅₀ was 42 µM. The nature of inhibition, though, mixed was for the most part competitive, since the competitive constant value (Ki = 7 µM) was lower than the non-competitive constant value (K′i = 35 µM).

In plasma membrane fractions, ATPase activity showed a high sensitivity to the heavy metal. It was non-competitively inhibited by cadmium in a narrow range of micromolar concentrations.