Project B01 – Molecular mechanisms of allosteric modulators at Y-receptors

This projects aims to identify the binding and activation mode of the peptide pancreatic polypeptide (PP) to the human Y4 receptor in the presence and absence of small molecule modulators. We will generate Y4R/Y1R chimeras to identify the binding region and will combine mutagenesis with 3D models of Y-receptors for the design of single mutants. Furthermore, by cell-free expression of the receptor, followed by photo-crosslinking of chemically modified peptides in combination with mass spectrometry the dynamics will be analyzed. As these compounds display high affinity at the same receptor, but differ in their activity, they will allow us to identify receptor residues involved in the different activities and finally to characterize different receptor states.

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 biochemie.lw.uni-leipzig.de

Resources

Unit Center Mass Spectrometry (>> Link)

  • Bruker Daltonics Microflex MALDI-ToF Massenspektrometer
  • Bruker Daltonics Ultraflex III MALDI-ToF/ToF
  • Bruker Daltonics Esquire HCT
  • Thermo Scientific Orbitrap Elite

Publications

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. https://doi.org/10.1038/s41596-020-00458-1.

Yang Z*, Han S*, Keller M*, Kaiser A*, Bender BJ*, Bosse M, Burkert K, Kögler LM, Wifling D, Bernhardt G, Plank N, Littmann T, Schmidt P, Yi C, Li B, Ye S, Zhang R, Xu B, Larhammar D, Stevens RC, Huster D, Meiler J, Zhao Q, Beck-Sickinger AG#, Buschauer A, Wu B#. Structural basis of ligand binding modes of neuropeptide Y Y1. Nature. 2018; 556:520-4.

Kögler LM, Stichel J, Kaiser A, Beck-Sickinger AG. Cell-Free Expression and Photo-Crosslinking of the Human Neuropeptide Y2 Receptor. Front Pharmacol. 2019; 10:176.

Wanka L, Babilon S, Kaiser A, Mörl K, Beck-Sickinger AG. Different mode of arrestin-3 binding at the human Y(1) and Y(2) receptor. Cell Signal. 2018; 50:58-71.

Schubert M, Stichel J, Du Y, Tough IR, Sliwoski G, Meiler J, Cox HM, Weaver CD, Beck-Sickinger AG. Identification and Characterization of the First Selective Y(4) Receptor Positive Allosteric Modulator. J Med Chem. 2017; 60:7605-12.

Burkert K*, Zellmann T*, Meier R, Kaiser A, Stichel J, Meiler J, Mittapalli GK, Roberts E, Beck-Sickinger AG. A Deep Hydrophobic Binding Cavity is the Main Interaction for Different Y(2) R Antagonists. ChemMedChem. 2017; 12:75-85.

Wanka L, Babilon S, Burkert K, Mörl K, Gurevich VV, Beck-Sickinger AG. C-Terminal motif of human neuropeptide Y(4) receptor determines internalization and arrestin recruitment. Cell Signal. 2017; 29:233-9.

Thieme V, Jolly N, Madsen AN, Bellmann-Sickert K, Schwartz TW, Holst B, Cox HM, Beck-Sickinger AG. High molecular weight PEGylation of human pancreatic polypeptide at position 22 improves stability and reduces food intake in mice. Br J Pharmacol. 2016; 173:3208-21.

Sliwoski G*, Schubert M*, Stichel J, Weaver D, Beck-Sickinger AG, Meiler J. Discovery of Small-Molecule Modulators of the Human Y4. PLoS One. 2016; 11:e0157146.

Mäde V, Babilon S, Jolly N, Wanka L, Bellmann-Sickert K, Diaz Gimenez LE, Mörl K, Cox HM, Gurevich VV, Beck-Sickinger AG. Peptide modifications differentially alter G protein-coupled receptor internalization and signaling bias. Angew Chem Int Ed Engl. 2014; 53:10067.

Pedragosa-Badia X, Sliwoski GR, Dong Nguyen E, Lindner D, Stichel J, Kaufmann KW, Meiler J, Beck-Sickinger AG. Pancreatic polypeptide is recognized by two hydrophobic domains of the human Y4 receptor binding pocket. J Biol Chem. 2014; 289:5846-59.