Covalent immobilization of recombinant fusion proteins with hAGT for single molecule force spectroscopy

Stefan K. Kufer, Hendrik Dietz, Christian Albrecht, Kerstin Blank, Angelika Kardinal, Matthias Rief, Hermann E. Gaub
2005 European Biophysics Journal  
A genetically modified form of the human DNA repair protein O 6 -alkylguanine-DNA-alkyltransferase (hAGT) was used to immobilize different recombinant hAGT fusion proteins covalently and selectively on gold and glass surfaces. Fusion proteins of hAGT with Glutathione S-Transferase and with tandem repeats of Titin Ig-domains, were produced and anchored via amino-polyethylene glycol benzylguanine. Anchoring was characterized and quantified with surface plasmon resonance, atomic force microscope
more » ... d fluorescence measurements. Individual fusion proteins were unfolded by single molecule force spectroscopy corroborating the selectivity of the covalent attachment.
doi:10.1007/s00249-005-0010-1 pmid:16160825 fatcat:afnh4iuzhfgj5esv7qlqiiaieu