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 taskforces and the training within IRTG.

Contact

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 uni-leipzig.de/agbs/

Dr. Patrick Scheerer (Project Leader)

Charité - Universitätsmedizin Berlin
Institute for Medical Physics and Biophysics
Group Structural Biology of Cellular Signaling
(Protein X-ray Crystallography - Cryo-Electron Microscopy - Signal Transduction)
Charitéplatz 1, D-10117 Berlin

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

Anja Koch (Technician)

Charité - Universitätsmedizin Berlin
Institute for Medical Physics and Biophysics
Group Structural Biology of Cellular Signaling
(Protein X-ray Crystallography - Cryo-Electron Microscopy - Signal Transduction)
Charitéplatz 1, D-10117 Berlin

Phone +49 30 450 524 150
E-Mail

Further Team Members

Dr. Michal Szczepek (Scientist in A01)
David Speck (PhD in A01)

Former Team Members

Brian Bauer (Technician)

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
Karin Mörl (Senior Scientist), Ronny Müller (Technician)

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

Biophysical methods – Charité Berlin
Michal Szczepek & Antje Kamprad (Scientist A01)

 

Publications

Haxhiraj D, Kuropka B, Brencher E, Rosenberger D, Brandt h, Speck D, Scheerer P, Brügger B, Clementi C, Freund C. T cell interaction partners of DHHC20. bioRxiv 2024.02.14.580290; doi: https://doi.org/10.1101/2024.02.14.580290&nbsp .

Sohail I, Laurin SA, Kleinau G, Chunila V , Kagiali ZCUK, Lohse MJ, Scheerer P, Bouvier M, McCormick P, Annibale P, Biebermann H. MRAP2 modifies the signaling and oligomerization state of the melanocortin-4 receptor. bioRxiv 2024.; doi: https://doi.org/10.1101/2024.04.09.588099.

Schmidt A, Kalms J, Lorent C, Katz S, Frielingsdorf S, Evans RM, Fritsch J, Siebert E, Teutloff C, Armstrong FA, Zebger I, Lenz O, Scheerer P. Stepwise conversion of the Cys6[4Fe-3S] to a Cys4[4Fe-4S] cluster and its impact on the oxygen tolerance of [NiFe]-hydrogenase. Chem Sci. 2023 Sep 20;14(40):11105-11120. doi: 10.1039/d3sc03739h. PMID: 37860641

Canzler S, Fischer M, Ulbricht D, Ristic N, Hildebrand PW, Staritzbichler R. ProteinPrompt: a webserver for predicting protein–protein interactions. Bioinformatics Advances. 2022, Volume 2, Issue 1, vbac059, https://doi.org/10.1093/bioadv/vbac059.

Nemec K, Schihada H, Kleinau G, Zabel U, Grushevskyi EO, Scheerer P, Lohse MJ, Maiellaro I. Functional modulation of PTH1R activation and
signaling by RAMP2. Proc Natl Acad Sci U S A. 2022; 119:e2122037119.

Höpfner F, Paisdzior S, Reininghaus N, Sohail I, Scheerer P, Annibale P, Biebermann H, Kühnen P. Evaluation of Pharmacological Rescue of Melanocortin-4 Receptor Nonsense Mutations by Aminoglycoside. Life (Basel). 2022; 12:1793.

Philippe A, Kleinau G, Gruner JJ, Wu S, Postpieszala D, Speck D, Heidecke H, Dowell SJ, Riemekasten G, Hildebrand PW, Kamhieh-Milz J, Catar R, Szczepek M, Dragun D, Scheerer P. Molecular Effects of Auto-Antibodies on Angiotensin II Type 1 Receptor Signaling and Cell Proliferation. Int J Mol Sci. 2022 Apr 2;23(7):3984. doi: 10.3390/ijms23073984. PMID: 35409344

Reininghaus N, Paisdzior S, Höpfner F, Jyrch S, Cetindag C, Scheerer P, Kühnen P, Biebermann H. A Setmelanotide-like Effect at MC4R Is Achieved by MC4R Dimer Separation. Biomolecules. 2022 Aug 15;12(8):1119. doi: 10.3390/biom12081119. PMID: 36009013.

Rudolf S, Kaempf K, Vu O, Meiler J, Beck-Sickinger AG, Coin I. Binding of natural peptide ligands to the neuropeptide Y5 receptor. Angew Chem
Int Ed Engl.
2022; 61:e202108738.

Speck D, Kleinau G, Meininghaus M, Erbe A, Einfeldt A, Szczepek M, Scheerer P, Pütter V. Expression and Characterization of Relaxin Family Peptide Receptor 1 Variants. Front Pharmacol. 2022 Jan 28;12:826112. doi: 10.3389/fphar.2021.826112. eCollection 2021. PMID: 35153771 Free PMC article.

Yang Y, Stensitzki T, Sauthof L, Schmidt A, Piwowarski P, Velazquez Escobar F, Michael N, Nguyen AD, Szczepek M, Brünig FN, Netz RR, Mroginski MA, Adam S, Bartl F, Schapiro I, Hildebrandt P, Scheerer P, Heyne K. Ultrafast proton-coupled isomerization in the phototransformation of phytochrome. Nat Chem. 2022 May 16. doi: 10.1038/s41557-022-00944-x. Online ahead of print. PMID: 3557791

Bergmann R, Chollet C, Els-Heindl S, Ullrich M, Berndt N, Pietzsch J, Máthé D, Bachmann M, Beck-Sickinger AG. Development of a ghrelin receptor inverse agonist for positron emission tomography. Oncotarget. 2021; 12:450-474.

Heyder NA, Kleinau G, Speck D, Schmidt A, Paisdzior S, Szczepek M, Bauer B, Koch A, Gallandi M, Kwiatkowski D, Bürger J, Mielke T, Beck-Sickinger A, Hildebrand P, Spahn CMT, Hilger D, Schacherl M, Biebermann H, Hilal T, Kühnen P, Kobilka BK, Scheerer P. Structures of active melanocortin-4 receptor−Gs-protein complexes with NDP-α-MSH and setmelanotide. Cell Research. 2021, 31(11):1176-1189. doi: 10.1038/s41422-021-00569-8. PMID: 34561620.

Işbilir A, Serfling R, Möller J, Thomas R, De Faveri C, Zabel U, Scarselli M, Beck-Sickinger AG, Bock A, Coin I, Lohse MJ, Annibale P. Determination of G-protein-coupled receptor oligomerization by molecular brightness analyses in single cells. Nat Protoc. 2021; 16:1419-1451.

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

Vogel A, Bosse M, Gauglitz M, Wistuba S, Schmidt P, Kaiser A, Gurevich VV, Beck-Sickinger AG, Hildebrand PW, Huster D. The dynamics of the neuropeptide Y receptor type 1 investigated by solid-state NMR and molecular dynamics simulation. Molecules. 2020; 25:5489.