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.
引用
收藏
页码：72 / 78
页数：6
相关论文
共 50 条
  • [1] Covalent immobilization of recombinant fusion proteins with hAGT for single molecule force spectroscopy
    Kufer, SK
    Dietz, H
    Albrecht, C
    Blank, K
    Kardinal, A
    Rief, M
    Gaub, HE
    EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, 2005, 35 (01): : 72 - 78
  • [2] Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
    Becke, Tanja D.
    Ness, Stefan
    Sudhop, Stefanie
    Gaub, Hermann E.
    Hilleringmann, Markus
    Schilling, Arndt F.
    Clausen-Schaumann, Hauke
    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2018, (138):
  • [3] Application of HaloTag Protein to Covalent Immobilization of Recombinant Proteins for Single Molecule Force Spectroscopy
    Taniguchi, Yukinori
    Kawakami, Masaru
    LANGMUIR, 2010, 26 (13) : 10433 - 10436
  • [4] Single-molecule force spectroscopy of proteins
    Rief, M
    Dietz, H
    Soft Condensed Matter Physics in Molecular and Cell Biology, 2006, : 249 - 258
  • [5] Single molecule force spectroscopy with individual proteins
    Fernandez, JM
    Gautel, M
    Rief, M
    Gaub, HE
    CURRENT CHALLENGES ON LARGE SUPRAMOLECULAR ASSEMBLIES, 1999, 519 : 319 - 336
  • [6] Covalent and selective immobilization of fusion proteins
    Kindermann, M
    George, N
    Johnsson, N
    Johnsson, K
    JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (26) : 7810 - 7811
  • [7] Covalent immobilization of single proteins on mica for molecular recognition force microscopy
    Klein, DCG
    Stroh, CM
    Jensenius, H
    van Es, M
    Kamruzzahan, ASM
    Stamouli, A
    Gruber, HJ
    Oosterkamp, TH
    Hinterdorfer, P
    CHEMPHYSCHEM, 2003, 4 (12) : 1367 - 1371
  • [8] SINGLE MOLECULE PROBING OF SNARE PROTEINS BY FORCE SPECTROSCOPY
    Parpura, V.
    JOURNAL OF NEUROCHEMISTRY, 2009, 108 : 55 - 55
  • [9] Single molecule force spectroscopy of modular proteins in the nervous system
    Fisher, TE
    Carrion-Vazquez, M
    Oberhauser, AF
    Li, HB
    Marszalek, PE
    Fernandez, JM
    NEURON, 2000, 27 (03) : 435 - 446
  • [10] A Lego Toolbox for Engineering Proteins for Single Molecule Force Spectroscopy
    Alonso-Caballero, Alvaro
    Rivas-Pardo, Jaime Andres
    Badilla, Carmen L.
    Echelman, Daniel J.
    Fernandez, Julio M.
    BIOPHYSICAL JOURNAL, 2017, 112 (03) : 300A - 300A
12345