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

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

2022

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.

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: 35577919.

2021

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.

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. 2021 Nov 25. doi: 10.1002/anie.202108738 . PMID:34822209.

2020

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

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

Contact

Dr. Patrick Scheerer (Project Leader)

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

Dr. Sylvia Els-Heindl (Scientist)

Brian Bauer (Technician)

Resources

Publications