Project Z03 – Peptide synthesis and membrane protein (GPCR) expression

The CRC consortium aims to investigate rhodopsin-like peptide-ligand GPCRs or adhesion GPCRs that are activated by peptide segments. The central goal of the Z03 project is to facilitate experimental approaches on the focused GPCRs by providing sufficient amounts of wild-type or chemically modified peptides as well as efficient GPCR-target plasmid construct design, test-expression and stabilization strategies of individual GPCR proteins for projects requiring to work in vitro on the purified samples. Furthermore, standardized approaches and protocols shall be worked out and applied. Z03 will be involved in the training within IRTG.

Contact

Dr. Patrick Scheerer (Project Leader)

Charité - Universitätsmedizin Berlin
Institute for Medical Physics and Biophysics
Group Protein X-ray Crystallography & Signal Transduction
Charitéplatz 1, D-10117 Berlin

Phone +49 30 450 524 178
E-Mail
Web biophysik.charite.de/forschung/ag_scheerer

Prof. Dr. Annette Beck-Sickinger (Project Leader)

Leipzig University, Faculty of Life Sciences
Institute of Biochemistry
Brüderstrasse 34, D-04103 Leipzig

Phone +49 341 97 36 900
E-Mail
Web biochemie.lw.uni-leipzig.de

Brian Bauer (Technician)

Charité - Universitätsmedizin Berlin
Institute for Medical Physics and Biophysics
Group Protein X-ray Crystallography & Signal Transduction
Charitéplatz 1, D-10117 Berlin

Phone +49 30 450 524 150
E-Mail

Resources

We can provide for the individual CRC-projects…

  • Individual peptide synthesis
    Strategic planning of in vitro GPCR experiments
  • Pre-testing of GPCR cloning/expression/purification/solubilisation
  • Production of G-protein or arrestin samples
  • Nanodisc/SMALPs integration for further investigations
  • Supervision, tips and tricks for cell culture techniques
  • Biophysical methods (in house) – MST, nDSF, BRET/FRET, SPR-like

Peptide Synthesis – Leipzig University
Sylvia Els-Heindl (Scientist), Ronny Müller (Technician)

Cloning/Expression/Protein purification/cell culture – Charité Berlin
Brain Bauer (Technician), David Speck (PhD), Anja Koch (Technician), Michal Szczepek (Scientist A01)

Biophysical methods – Charité Berlin
Michal Szczepek (Scientist A01)

Publications

Paisdzior S, Dimitriou IM, Schöpe PC, Annibale P, Scheerer P, Krude H, Lohse MJ, Biebermann H, Kühnen P. Differential Signaling Profiles of MC4R Mutations with Three Different Ligands. Int J Mol Sci. 2020 Feb 12;21(4). pii: E1224. doi: 10.3390/ijms21041224. PMID: 32059383