Details

Autor(en) / Beteiligte

• Hainfeld, James F.
• Liu, Wenqiu
• Halsey, Carol M.R.
• Freimuth, Paul
• Powell, Richard D.

Titel

Ni–NTA–Gold Clusters Target His-Tagged Proteins

Ist Teil von

• Journal of structural biology, 1999-09, Vol.127 (2), p.185-198

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

1999

Quelle

Access via ScienceDirect (Elsevier)

Beschreibungen/Notizen

• 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.