Ni-NTA-gold clusters target His-tagged proteins

Addition of six histidines to recombinant proteins has proved useful in their purification by nickel-affinity columns. This technology was adapted by synthesizing the chelator for nickel (nitrilotriacetic acid, NTA) onto the surface of gold clusters. These Ni-NTA-gold clusters were shown to specifically target the 6His region of tagged proteins. Results were verified by column chromatography, dot and overlay blots, UV-Vis spectroscopy, and scanning transmission electron microscopy. A 6His-tagged adenovirus "knob" protein was also shown to maintain receptor binding activity after gold labeling. Two types of gold clusters were used: 1.4-nm Nanogold and a new 1.8-nm "PeptideGold" coated with an NTA-dipeptide-thiol. These novel labels should be useful in site-specific high-resolution EM labeling, as well as in metallographic development, detection in the light microscope, or direct visualization.