Publication details
Electrochemical investigation of metallothionein and apo-metallothionein
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Proceedings |
| Conference | 11th International conference on electroanalysis |
| MU Faculty or unit | |
| Citation | |
| Field | Electrochemistry |
| Keywords | Metallothionein; apo-metallothionein; biosensor; electrochemistry; carcinogenesis; spontaneous mutagenesis; tumour; |
| Description | Introduction. Metallothionein (MT) belongs to group of intracellular, cysteine-rich proteins with molecular weight from 6 to 10 kDa, which are usually abundant at low content at human organisms. Their marked expression could be caused by many exogenous and endogenous factors. During last 15 years number of authors are has been concerning with study of MT involvement in carcinogenesis and spontaneous mutagenesis, because an increasing amount of MT in tumour cells probably relates with cell proliferation. On the base of the abovementioned reasons, it is necessary to develop a method that could be used not only for determination of MT but also for study of MT structural changes during stress conditions. Objective. The aim of this work was to study the effect of ethylendiamine-N, N, N`, N`-tetraacetic acid (EDTA) on MT structure by means of electrochemical techniques. Materials and methods. Electrochemical measurements were performed with AUTOLAB Analyser connected to VA-Stand 663, using a standard cell with three electrodes. The working electrode was a hanging mercury drop electrode. The reference electrode was Ag/AgCl/3M KCl electrode and the auxiliary electrode was the graphite electrode. The samples of the MT were reduced before each measurement. Results. It is known that EDTA is able to bind heavy metals not only from surrounding medium but also from structures of biological compounds. Here we added the different EDTA concentrations (up to 500 nM) to 100 nM MTCd5Zn2. The sample was analysed by adsorptive transfer stripping differential pulse voltammetry. On the base of the obtained results it follows that the signals, which correspond to reduction of zinc and cadmium present in MT structure, disappeared with increasing EDTA concentration. In addition we obtained the apo-MT (MT without heavy metal) at the highest concentration of EDTA (500 nM). The comparison between MT and apo-MT signals have been done and discussed. Conclusions. The described technique offers a good tool not only for study of heavy metal amounts in MT structure but also for characterization of conformation changes in MT structure. |
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