Bioanalytical Method to Determine the Effects of Cyanide, Cyanide Metabolites and Cyanide Antidotes on the Activity of Cytochrome C Oxidase Immobilized in an Electrode Supported Lipid Bilayer Membrane

摘要

The bioanalytical studies pursued in this research take advantage of the electron transfer reactivity of cytochrome c oxidase (CCO) when immobilized in a bilayer attached to an electrode surface. This enzyme immobilization method enables CCO to exhibit reaction chemistry that mimics its in vivo behavior because the bilayer structure is thought to reproduce its in vivo environment in the inner mitochondrial membrane. In addition this experimental platform for CCO enables an innovative and direct screening assay for toxic substances and potential antidotes for inhibition of this enzyme by these substances. The preliminary results obtained thus far describe the following: 1) An alternate programmable reference oscillator that is compatible with the electronic circuitry in the quartz crystal microbalance obtained from CH Instruments. 2) Octadecyl mercaptan deposition control. 3) Possibility for chloride interference in the potential window of the experiment. 4) Additional analytical methods to supplement electrochemical results obtained. a. Spectroelectrochemistry experimental design. b. New bioanalytical method for detecting cyanide using metmyoglobin.