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Expression and purification of recombinant proteins

Various biophysical methods require large amounts of recombinantly expressed and purified proteins. Additionally, preparation of stable isotope labeled protein is a prerequisite for elucidating their three-dimensional structures by NMR. Therefore, we construct and use efficient recombinant expression systems for the proteins of our interest.

  • Bleffert F, Granzin J, Gohlke H, Batra-Safferling R, Jaeger KE, Kovacic F 
    Pseudomonas aeruginosa esterase PA2949, a bacterial homolog of the human membrane esterase ABHD6: expression, purification and crystallization. 
    Acta Crystallography F75, 270-277 (2019) 
    https://doi.org/10.1107/S2053230X19002152  
    http://scripts.iucr.org/cgi-bin/paper?S2053230X19002152  
  • Klinker S, Stindt S, Gremer L, Bode JG, Gertzen CGW, Gohlke H, Weiergräber OH, Hoffmann S, Willbold D 
    Phosphorylated tyrosine 93 of hepatitis C virus nonstructural protein 5A is essential for interaction with host c-Src and efficient viral replication 
    J Biol Chem 294, 7388-7402 (2019) 
    https://doi.org/10.1074/jbc.RA119.007656  
  • Kukuk L, Dingley AJ, Granzin J, Nagel-Steger L, Thiagarajan-RosenkranzP, Ciupka D, Hänel K, Batra-Safferling R, Pacheco V, Stoldt M, PfefferK, Beer-Hammer S, Willbold D, Koenig BW 
    Structure of the SLy1 SAM homodimer reveals a new interface for SAM domain self-association 
    Scientific Reports 9, 54 (2019) 
    https://doi.org/10.1038/s41598-018-37185-3  
  • Viennet T, Bungert-Pümke S, Elter S, Viegas A, Fahlke C, Etzkorn M 
    Reconstitution and NMR characterization of the ion-channel accessory subunit barttin in detergents and lipid-bilayer nanodiscs 
    Frontiers in Molecular Biosciences 6, 13 (2019) 
    https://doi.org/10.3389/fmolb.2019.00013  
  • Santur K, Sevenich M, Schwarten M, Nischwitz V, Willbold D, Mohrlüder J 
    In Vitro Reconstitution of the Highly Active and Natively Folded Recombinant Human Superoxide Dismutase 1 Holoenzyme 
    ChemistrySelect 3, 7627-7632 (2018) 
    https://doi.org/10.1002/slct.201801319  
    https://onlinelibrary.wiley.com/doi/abs/10.1002/slct.201801319  
  • Dammers C, Reiss K, Gremer L, Lecher J, Ziehm T, Stoldt M, Schwarten M, Willbold D.  
    Pyroglutamate-modified amyloid-β(3-42) shows α-helical intermediates before amyloid formation. 
    Biophys. J. 112, 1621-1633 (2017) 
  • Bronder AM, Bieker A, Elter S, Etzkorn M, Haeussinger D, Oesterhelt F 
    Oriented membrane protein reconstitution into tethered lipid membranes for AFM Force Spectroscopy 
    Biophys. J. 111, 1925-1934 (2016) 
    http://dx.doi.org/10.1016/j.bpj.2016.08.051  
  • Hänel K, Möckel L, Brummel M, Peiris K, Hartmann R, Dingley AJ, Willbold D, Loidl-Stahlhofen A 
    Expression and purification of soluble HIV-2 viral protein R (Vpr) using a sandwich-fusion protein strategy 
    Prot. Expr. Purific. 95, 156-161 (2014) 
    http://www.sciencedirect.com/science/article/pii/S1046592813002763  
  • Mirecka EA, Gremer L, Schiefer S, Oesterhelt F, Stoldt M, Willbold D, Hoyer W 
    Engineered aggregation inhibitor fusion for production of highly amyloidogenic human islet amyloid polypeptide 
    J. Biotechnol. 191, 221-227 (2014) 
    http://dx.doi.org/10.1016/j.jbiotec.2014.06.006  
  • Marbach, J; Zentis, P; Ellinger, P; Müller, H; Birkmann, E 
    Expression and characterization of fully posttranslational modified cellular prion protein in Pichia pastoris 
    Biol. Chem. 394, 1475-1483 (2013) 
  • Glück JM 
    Ligand interaction analysis of membrane-anchored proteins. 
    Schriften des Forschungszentrums Jülich Reihe Gesundheit, Band 31, (2011) 
    http://wwwzb1.fz-juelich.de/contentenrichment/inhaltsverzeichnisse/2010/verlag/gesund31_i.pdf  
  • Glück JM, Hoffmann S, Koenig BW, Willbold D 
    Single Vector System for Efficient N-myristoylation of Recombinant Proteins in E. coli 
    PLoS ONE 5, e10081 (2010) 
    full text open access  
    The plasmids in this publication can be received via https://www.addgene.org/Dieter_Willbold/ 
  • Wiesehan K, Funke SA, Fries M, Willbold D 
    Purification of recombinantly expressed and cytotoxic human amyloid-beta peptide. 
    J. Chrom. B 856, 229-233 (2007) 
  • Wittlich M, Wiesehan K, Koenig BW, Willbold D 
    Expression, purification, and membrane reconstitution of a CD4 fragment comprising the transmembrane and cytoplasmic domains of the receptor. 
    Protein Expr. Purif. 55, 198-207 (2007) 
  • Koenig BW, Rogowski M, Louis JM 
    A rapid method to attain isotope labeled small soluble peptides for NMR studies. 
    J. Biomol. NMR 26, 193-202 (2003) 
  • Preusser A, Jonas G and Willbold D 
    Purification of recombinantly expressed human cluster determinant 4 cytoplasmatic domain. 
    J. Chromatogr. B 786, 39-44 (2003) 
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