Application of electroanalytical methods to the characterization of metallothioneins and related molecules

Abstract

Electroanalytical methods have been applied to the characterization and to the study of complexing properties of several Cd,Zn metallothioneins (MT) of different mammalian origin, rabbit and rat liver and horse kidney and human foetal liver and adult kidney. Differential pulse polarography (DPP) has been selected as the most suitable technique for distinguishing the chemical form of compounds and, assuming a diffusion controlled mechanism, monitoring the evolution of different species as a function of parameters, such as the solution pH and the gradual addition of cations, cadmium and/or zinc, initially contained in the studied molecules. Due to the fact that the metallothioneins structure is complex and often not perfectly known, the peptidic fragment Lys-Cys-Thr-Cys-Cys-Ala (56-61) MT-1 (FT) of the mouse liver has been taken as a model for MT characterization. The study of FT alone and in the presence of Cd and/or Zn, using several electroanalytical methods has been very useful for the investigation of adsorption-diffusion processes and for the identification and elucidation of electrochemical systems. The interpretation of voltammetric data is very often not easy. For some cases, proper interpretation can be made using chemometric techniques. Multivariate factor analysis has been applied to the electroanalytical investigation of the complexing properties between cadmium and/or zinc and FT, using results obtained by DPP and linear sweep voltammetry. For more complex systems, like MT, the application of chemometry becomes more complex.