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

被引：0
|
作者：
Stefan K. Kufer
Hendrik Dietz
Christian Albrecht
Kerstin Blank
Angelika Kardinal
Matthias Rief
Hermann E. Gaub
机构：
[1] Ludwig-Maximilians-Universität München and Center for NanoScience,Lehrstuhl für Angewandte Physik, Sektion Physik
[2] Technische Universität München,Physik
来源：
European Biophysics Journal | 2005年 / 35卷
关键词：
Molecular recognition ; SPR ; AFM ; Suicide coupler ; hAGT ; SNAP-tag;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
A genetically modified form of the human DNA repair protein O6-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.
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页码：72 / 78
页数：6
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