CRUI: Quantification of Metal Transfer From Metallothionein to Apometalloproteins Using Voltammetry Combined On Line with HPLC/ICP-MS

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Life has evolved in close association with metals and it is not surprising, therefore, that evolutionary processes have selectively incorporated specific metals to take advantage of the additional chemical attributes that these metals bring to biological molecules. Thus, the incorporation of Zn or Cu metal may confer additional steric possibilities, catalytic activity or redox properties to the acceptor molecule and these metalloproteins are known to control important biological processes as varied as gene expression, respiration and acid/base balance.One intracellular protein that has been advocated to be involved in all of these processes in metallothionein, although the mechanisms by which this protein controls cellular metal homeostasis is unclear. A novel technique (EC/HPLC/ICP-MS) involving directly coupling electrochemistry (EC), high performance liquid chromatography (HPLC) and inductively coupled plasma mass spectroscopy (ICP-MS) will be developed to conduct a series of experiments that have been collectively designed to determine (i) the relative importance of glutathione and redox conditions necessary to cause the donation and acceptance of essential metals between metallothionein and apoenzymes within a complex cytoplasmic milieu; (ii) whether glutathione disulfide or oxidizing conditions can lead to the release of Cd and other physiologically toxic metals from metallothionein causing subsequent non-specific binding to target macromolecules.