Jump to contentJump to search

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.

  • Brown H, Chung M, Üffing A, Batistatou N, Tsang T, Doskocil S, Mao W, Willbold D, Bast RC Jr, Lu Z, Weiergräber OH, Kritzer JA 
    Structure-based design of stapled peptides that bind GABARAP and inhibit autophagy 
    J Am Chem Soc 144, 14687-14697 (2022) 
    https://doi.org/10.1021/jacs.2c04699  
  • Huwa N, Weiergräber OH, Fejzagić AV, Kirsch C, Schaffrath U, Classen T 
    The crystal structure of the defense conferring rice protein OsJAC1 reveals a carbohydrate binding site on the dirigent-like domain 
    Biomolecules 12, 1126 (2022) 
    https://doi.org/10.3390/biom12081126  
  • Cukkemane A, Becker N, Zielinski M, Frieg B, Lakomek NA, Heise H, SchröderGF, Willbold D, Weiergräber OH 
    Conformational heterogeneity coupled with β-fibril formation of a scaffold protein involved in chronic mental illnesses 
    Transl Psychiatry 11, 639 (2021) 
    https://doi.org/10.1038/s41398-021-01765-1  
  • Freischem S, Grimm I, López-Pérez A, Willbold D, Klenke B, Vuong C, DingleyAJ, Weiergräber OH 
    Interaction mode of the novel monobactam AIC499 targeting penicillin binding protein 3 of Gram-negative bacteria 
    Biomolecules 11, 1057 (2021) 
    https://doi.org/10.3390/biom11071057  
  • Huwa N, Weiergräber OH, Kirsch C, Schaffrath U, Classen T 
    Biochemical and initial structural characterization of the monocot chimeric jacalin OsJAC1 
    Int J Mol Sci 22, 5639 (2021) 
    https://doi.org/10.3390/ijms22115639  
  • López-Pérez A, Freischem S, Grimm I, Weiergräber O, Dingley AJ, López-AlbercaMP, Waldmann H, Vollmer W, Kumar K, Vuong C 
    Discovery of pyrrolidine-2,3-diones as novel inhibitors of P. aeruginosa PBP3 
    Antibiotics 10, 529 (2021) 
    https://doi.org/10.3390/antibiotics10050529  
  • 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) 
Responsible for the content: