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Eur Biophys J. 2005 Dec;35(1):72-8. Epub 2005 Sep 14.

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

Author information

1
Lehrstuhl für Angewandte Physik, Sektion Physik, Ludwig-Maximilians-Universität München and Center for NanoScience, Amalienstrasse 54, 80799 Munich, Germany.

Abstract

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 and fluorescence measurements. Individual fusion proteins were unfolded by single molecule force spectroscopy corroborating the selectivity of the covalent attachment.

PMID:
16160825
DOI:
10.1007/s00249-005-0010-1
[Indexed for MEDLINE]

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