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AKB: Biologische Physik

AKB 200: Poster Session II

AKB 200.36: Poster

Dienstag, 8. März 2005, 17:00–19:00, Poster TU C

Covalent immobilisation of recombinant fusion proteins with hAGT for single molecule force spectroscopy — •Stefan Kufer and Hermann E. Gaub — Amalienstr.54(I); 80799 München

A genetically modified form of the human DNA repair protein O6-alkylguanine-DNA-alkyltransferase (hAGT) was used to immobilize different recombinant hAGT fusion proteins covalent and selective on gold and glass surfaces. Fusion proteins of hAGT with Glutathione S-Transferase (GST) and with tandem repeats of Titin Ig domains, were produced and anchored via amino-polyethylene glycol (PEG)-benzylguanine (BG). Anchoring was characterized and quantified with surface plasmon resonance (SPR), atomic force microscope (AFM) and fluorescence measurements. Individual fusion proteins were unfolded by single molecule force spectroscopy corroborating the selectivity of the covalent attachment.