Copper proteins immobilised on gold electrodes for (bio)analytical studies

Copper electrochemistry at modified gold electrodes was investigated with two different states of the metal ion: first bound in azurin from Pseudomonas aeruginosa and second introduced via metal ion uptake in metallothionein (NIT) from rabbit liver. Azurin was immobilised on a mercaptosuccinic acid (MSA) layer self-assembled on gold. The redox behaviour in the adsorbed as well as in the covalently immobilised state was found to be quasi-reversible with a formal potential of + 198 mV versus Ag/AgCl. The pH variation suggests an optimal pH range for efficient electrode communication in the neutral range. NIT was fixed at electrochemically cleaned gold using the accessible cysteins of the protein. Copper was found to bind to the MT-modified gold electrode. The electrochemical behaviour of the bound copper was characterised in copper-free solution with a formal potential of + 245 mV versus Ag/AgCl. Stability and potential use is discussed. (C) 2002 Published by Elsevier Science B.V.