Abstract
Isothermal titration calorimetry (ITC) was used to study the binding of Cd2+ and Zn2+ by glutathione (GSH) and phytochelatins (PCn), the metal sequestering compounds in plants and algae. The results are compared with those obtained by differential pulse polarography (DPP) assisted by multivariate curve resolution with alternating least squares (MCR–ALS) and by electrospray ionization mass spectrometry (ESI–MS). ITC allows one to determine (i) the stoichiometries of the different complexes (and confirms those found by DPP/MCR–ALS and ESI–MS) and (ii) their binding and thermodynamic parameters. For Cd–PC4, the sequential binding sites model with two identical sites yields the best fitting of ITC curves and confirms the presence of CdPC4 and Cd2PC4 complexes. For Zn–PC4, exothermic formation of ZnPC4 is reported. Conditional stability and formation constants for Cd–GSH and Zn–GSH are determined from the fitting of the proper model to experimental ITC curves. The effect of different buffers in the complexation processes shows the key role of the choice of the buffer in calorimetric study.
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