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

doi:10.1007/s00249-005-0010-1

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

University of Munich

Research output: Contribution to journal › Article › peer-review

43 Scopus citations

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.

Original language	English
Pages (from-to)	72-78
Number of pages	7
Journal	European Biophysics Journal
Volume	35
Issue number	1
DOIs	https://doi.org/10.1007/s00249-005-0010-1
State	Published - Dec 2005

Keywords

AFM
Molecular recognition
SNAP-tag
SPR
Suicide coupler
hAGT

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1007/s00249-005-0010-1

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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.",

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AU - Kufer, Stefan K.

AU - Dietz, Hendrik

AU - Albrecht, Christian

AU - Blank, Kerstin

AU - Kardinal, Angelika

AU - Rief, Matthias

AU - Gaub, Hermann E.

PY - 2005/12

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N2 - 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.

AB - 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.

KW - AFM

KW - Molecular recognition

KW - SNAP-tag

KW - SPR

KW - Suicide coupler

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

Abstract

Keywords

UN SDGs

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