Cadmium decreases the levels of glutathione and enhances the phytochelatin concentration in the marine dinoflagellate Lingulodinium polyedrum

The toxic effects of metals on the aquatic environment are documented and well known. Metals are able to unbalance the intracellular redox potential and, therefore, induce the oxidative stress in living organisms. In this study, the responses of glutathione [reduced (GSH) and oxidized (GSSG)] and the structurally GSH-related peptides phytochelatins 3 and 4 (PC3 and PC4) were examined in the marine dinoflagellate Lingulodinium polyedrum exposed to cadmium (Cd). A novel method for PC3 and PC4 synthesis is described here based on a Boc strategy, yielding peptides with purities higher than 97 %. Analytical detection of GSH, GSSG, PC3, and PC4 applying liquid chromatography-tandem mass spectrometry (LC-MS/MS) method had been developed and described as robust and accurate with a detection limit of nmol g(-1) dry weight (DW) for PC3 and PC4. The intracellular levels of GSH and GSSG decreased dramatically over 24 and 48-h exposure to 18 mu mol L-1 Cd. These decreases were followed by the enhancement of intracellular PC3 and PC4 levels, in which syntheses started to be detected after 2 and 8-h exposure, respectively. Moreover, the PC4/PC3 ratio reached its maximum over 24-h exposure, being 8 to 75-fold higher than the one observed for other microalgae. This seems to be an efficient strategy of L. polyedrum to be protected against Cd environment contamination, since PC4 has more chelating sites and is structurally more stable than PC2 and PC3, the most abundant for other microalgae.