Gold nanocluster formation using metallothionein: mass spectrometry and electron microscopy.
Clonable contrasting agents for light microscopy, such as green fluorescent protein, have revolutionized biology, but few such agents have been developed for transmission electron microscopy (TEM). As an attempt to develop a novel clonable contrasting agent for TEM, we have evaluated metallothionein, a small metal-binding protein, reacted with aurothiomalate, an anti-arthritic gold compound. Electro spray ionization and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry measurements show a distribution of gold atoms bound to individual metallothionein molecules. Unlike previous reports, these data show gold binding occurred as the addition of single atoms without retention of additional ligands. Moreover, under certain conditions, MALDI spectra show gold binding ratios of greater than 1:1 with the cysteine residues of metallothionein. Together, this may hint at a gold-binding mechanism similar to gold nanocluster formation. Finally, metallothionein-gold complexes visualized in the TEM show a range of sizes similar to those used as current TEM labels, and show the potential of the protein as a clonable TEM label in which the gold cluster is grown on the label, thereby circumventing the problems associated with attaching gold clusters.
Mercogliano
Christopher P
CP
Department of Biology, Brandeis University, 415 South Street, Waltham, MA 02454, USA.
DeRosier
David J
DJ
D006046:Gold / Q000737:chemistry*; D006052:Gold Sodium Thiomalate / Q000737:chemistry; D007202:Indicators and Reagents; D008024:Ligands; D008668:Metallothionein / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D049329:Nanostructures / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling
D007202:Indicators and Reagents; D008024:Ligands; D006052:Gold Sodium Thiomalate; D006046:Gold; D008668:Metallothionein