Creation of a gold nanoparticle based electrochemical assay for the detection of inhibitors of bacterial cytochrome bd oxidases.
Cytochrome bd oxidases are membrane proteins expressed by bacteria including a number of pathogens, which make them an attractive target for the discovery of new antibiotics. An electrochemical assay is developed to study the activity of these proteins and inhibition by quinone binding site tool compounds. The setup relies on their immobilization at electrodes specifically modified with gold nanoparticles, which allows achieving a direct electron transfer to/from the heme cofactors of this large enzyme. After optimization of the protein coverages, the assay shows at pH7 a good reproducibility and readout stability over time, and it is thus suitable for further screening of small molecule collections.
Laboratoire de Bioelectrochimie et Spectroscopie, UMR 7140, Chimie de la Matière Complexe, Université de Strasbourg, CNRS, Strasbourg, France.
Eugénie
E
Fournier
Laboratoire de Bioelectrochimie et Spectroscopie, UMR 7140, Chimie de la Matière Complexe, Université de Strasbourg, CNRS, Strasbourg, France.
Anton
A
Nikolaev
Johann Wolfgang Goethe-University Frankfurt, Max-von-Laue-Straße 7, D-60438 Frankfurt am Main, Germany.
Hamid R
HR
Nasiri
Albert-Ludwigs-Universität Freiburg, Institut für Biochemie, Freiburg, Germany.
Jo
J
Hoeser
Albert-Ludwigs-Universität Freiburg, Institut für Biochemie, Freiburg, Germany.
Thorsten
T
Friedrich
Laboratoire de Bioelectrochimie et Spectroscopie, UMR 7140, Chimie de la Matière Complexe, Université de Strasbourg, CNRS, Strasbourg, France. Electronic address: hellwig@unistra.fr.
Petra
P
Hellwig
Laboratoire de Bioelectrochimie et Spectroscopie, UMR 7140, Chimie de la Matière Complexe, Université de Strasbourg, CNRS, Strasbourg, France. Electronic address: fmelin@unistra.fr.
Frederic
F
Melin
D003573:Cytochrome b Group; D003580:Cytochromes / Q000037:antagonists & inhibitors*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004563:Electrochemistry / Q000379:methods*; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors*; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*
D003573:Cytochrome b Group; D003580:Cytochromes; D045222:Electron Transport Chain Complex Proteins; D004791:Enzyme Inhibitors; D029968:Escherichia coli Proteins; D006046:Gold; D010088:Oxidoreductases; C040077:cytochrome bd terminal oxidase complex, E coli
Antibiotics; Functional assay; Gold nanoparticles; Inhibitors; Membrane proteins; Terminal oxidases