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.
- Amariei DA, Pozhydaieva N, David B, Schneider P, Classen T, Gohlke H, Weiergräber OH, Pietruszka J
Enzymatic C3-Methylation of Indoles Using Methyltransferase PsmD─Crystal Structure, Catalytic Mechanism, and Preparative Applications
ACS Catal 12, 14130-14139 (2022)
https://doi.org/10.1021/acscatal.2c04240
- 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
- Weiergräber OH, Petrović D, Kislat A, Pattky M, Fabig J, Batra-Safferling R, Schulte am Esch J, Hänel K, Huhn C, Strodel B, Homey B, Willbold D
Structure and Dynamics of Human Chemokine CCL16—Implications for Biological Activity
Biomolecules 12, 1588 (2022)
https://doi.org/10.3390/biom12111588
- 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)
- Schlesinger R, Cousin A, Granzin J, Batra-Safferling R.
Expression and purification of arrestin in yeast Saccharomyces cerevisiae.
Methods Cell Biol 142, 159-172 (2017)
https://doi.org/10.1016/bs.mcb.2017.07.003
- 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)