title: Microbial metalloproteomes explored using MIRAGE.
abstract: Metalloproteomics is a rapidly developing field of science that involves the comprehensive analysis of all metal-containing or metal-binding proteins in a biological sample. The purpose of this review is to offer an overview of the research involving Metal Isotope native RadioAutography in Gel Electrophoresis (MIRAGE), a powerful new method to visualize and study the proteome of a particular metal ion. MIRAGE involves four steps: i) labelling of target proteins with a radioisotope; ii) separation of intact holo-proteins using native isoelectric focusing (1D) combined with Blue Native PAGE (2D); iii) spot visualization and quantification using autoradiography; and iv) protein identification by tandem mass spectrometry. MIRAGE Investigations of the soluble Cu, Zn, and Fe metalloproteomes of Escherichia coli, and of the soluble Mo and W proteomes of the hyperthermophilic archaeon Pyrococcus furiosus are reviewed.
authors.affiliation: Department of Biotechnology, Delft University of Technology, Julianalaan 67, NL-2628 BC Delft.
authors.forename: Ana-Maria
authors.initials: AM
authors.lastname: Sevcenco
authors.forename: Wilfred R
authors.initials: WR
authors.lastname: Hagen
authors.forename: Peter-Leon
authors.initials: PL
authors.lastname: Hagedoorn
mesh_terms: D001345:Autoradiography; D001419:Bacteria / Q000235:genetics*; D008667:Metalloproteins / Q000737:chemistry*; D008827:Microbiological Phenomena; D063347:Native Polyacrylamide Gel Electrophoresis; D020543:Proteome / Q000235:genetics*; D053719:Tandem Mass Spectrometry
chemical_list: D008667:Metalloproteins; D020543:Proteome