Project A04 – Characterizing the molecular interaction between the Y receptor and arrestin
We aim at providing structural data on the active complex of Y receptors with arrestin as well as information on the modification of the equilibrium dynamics of the receptor. To achieve this, we will (i) combine peptide synthesis and protein ligation techniques to prepare NMR-active proteins with a few isolated isotopic labels and EPR-active proteins. (ii) We will use solution and solid-state NMR techniques to determine the backbone structure of segments of arrestin in interaction with Y receptors. (iii) We will determine structural constraints to determine the mutual orientation and will verify this with mutagenesis of both, arrestin and Y receptors. (iv) We will investigate the molecular dynamics of the Y receptors in response to agonist and arrestin binding as well as dynamic alterations in response to conformational changes arrestin undergoes upon binding to the receptor.
Contact
Prof. Dr. Daniel Huster (Project Leader)
Leipzig University, Faculty of Medicine
Institute of Medical Physics and Biophysics
Härtelstraße 16 – 18, 04107 Leipzig
Phone +49 341 97 15701
E-Mail
Web biophysik.medizin.uni-leipzig.de
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 36900
E-Mail
Web biochemie.lw.uni-leipzig.de
Resources
- EPR-spectrometer
- NMR techniques
Publications
Dey S, Surendran D, Engberg O, Gupta A, Fanibunda SE, Das A, Maity BK, Dey A, Visvakarma V, Kallianpur M, Scheidt HA, Walker G, Vaidya VA, Huster D, Maiti S. Altered Membrane Mechanics Provides a Receptor-Independent Pathway for Serotonin Action. Chemistry. 2021 May 12;27(27):7533-7541. doi: 10.1002/chem.202100328. Epub 2021 Mar 12. PMID: 33502812.
Vu O, Bender BJ, Pankewitz L, Huster D, Beck-Sickinger AG, Meiler J. The Structural Basis of Peptide Binding at Class A G Protein-Coupled Receptors. Molecules. 2021 Dec 30;27(1):210. doi:10.3390/molecules27010210. PMID: 35011444 Review.
Krug U, Gloge A, Schmidt P, Becker-Baldus J, Bernhard F, Kaiser A, Montag C, Gauglitz M, Vishnivetskiy SA, Gurevich VV, Beck-Sickinger AG, Glaubitz C, Huster D. The Conformational Equilibrium of the Neuropeptide Y2 Receptor in Bilayer Membranes. Angew Chem Int Ed Engl. 2020 Aug 13. doi: 10.1002/anie.202006075. Epub ahead of print. PMID: 32790043
Laugwitz JM, Haeri HH, Kaiser A, Krug U, Hinderberger D, Beck-Sickinger AG, Schmidt P. Probing the Y2 Receptor on Transmembrane, Intra- and Extra-Cellular Sites for EPR Measurements. molecules. Accepted: 09092020.
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 receptor. Nature. 2018; 556:520-4.
Schmidt P, Bender BJ, Kaiser A, Gulati K, Scheidt HA, Hamm HE, Beck-Sickinger AG, Meiler J, Huster D. Improved in vitro folding of the Y2 G protein-coupled receptor into bicelles. Front Plant Sci. 2018; 4:100.
Zernia S, Ott F, Bellmann-Sickert K, Frank R, Klenner M, Jahnke HG, Prager A, Abel B, Robitzki A, Beck-Sickinger AG. Peptide-mediated specific immobilization of catalytically active cytochrome P450 BM3 variant. Bioconjug Chem. 2016; 27:1090-7.
Kaiser A, Müller P, Zellmann T, Scheidt HA, Bosse M, Meier R, Meiler J, Huster D, Beck-Sickinger AG, Schmidt P. NMR-guided structural model of neuropeptide Y Bound to its G protein-coupled Y2. Angew Chem Int Ed. 2015; 52:7446-9.
Gimenez LE, Babilon S, Wanka L, Beck-Sickinger AG, Gurevich VV. Mutations in arrestin-3 differentially affect binding to neuropeptide Y receptor subtypes. Cell Signal. 2014; 26:1523-31.
Schmidt P, Scheidt HA, Thomas L, Müller P, Huster D. The G protein-coupled neuropeptide Y receptor Type 2 is highly dynamic in lipid membranes. Chemistry. 2014; 20:4986-92.
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.
Berger C, Berndt S, Pichert A, Theisgen S, Huster D. Efficient isotopic tryptophan labeling of membrane proteins by an indole controlled process conduct. Biotechnol Bioeng. 2013; 110:1681-90.
Nordsieck K, Pichert A, Samsonov SA, Thomas L, Berger C, Pisabarro MT, Huster D, Beck-Sickinger AG. Residue 75 of interleukin-8 is crucial for its interactions with glycosaminoglycans. ChemBioChem. 2012; 13:2558-66.
Bosse M, Thomas L, Hassert R, Beck–Sickinger AG, Huster D, Schmidt P. Assessment of a fully active class A GPCR isolated from in vitro. Biochemistry. 2011; 50:9817-25.
