Project A04 – Characterizing the molecular interaction between the Y receptor and arrestin
We investigate the interaction between neuropeptide Y receptors (YRs) and arrestin with the focus on dynamic aspects of the receptors and their interaction with arrestin in the biological membrane environment. To achieve this, we will (i) combine peptide synthesis and protein ligation techniques to prepare NMR-active proteins with 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 YRs. (iii) We will analyze the dynamics of the interaction and verify essential interaction partners with mutagenesis of both arrestin and YRs. (iv) Furthermore, we will investigate the molecular dynamics of the YRs in response to agonist and arrestin binding as well as dynamic alterations in response to conformational changes arrestin undergoes upon binding to the receptor.
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 uni-leipzig.de/agbs/
Matthias Voitel (PhD-Student)
Leipzig University, Faculty of Medicine
Institute of Medical Physics and Biophysics
Härtelstrasse 16 – 18, 04107 Leipzig
Varsha Gupta (PhD-Student)
Leipzig University, Faculty of Life Sciences
Institute of Biochemistry
Brüderstrasse 34, D-04103 Leipzig
Resources
- EPR-spectrometer
- Bruker Avance Neo 700 MHz NMR spectrometer
Publications
Coin I, Huster D. Ready for the sheet: β-strand folding of phosphorylation clusters guides GPCR binding to arrestin. Structure. 2023 Nov 2;31(11):1289-1291. doi: 10.1016/j.str.2023.09.013 . PMID: 37922864 .
Gupta A, Krupa P, Engberg O, Krupa M, Chaudhary A, Li MS, Huster D, Maiti S. Unusual Robustness of Neurotransmitter Vesicle Membranes against Serotonin-Induced Perturbations. J Phys Chem B. 2023 Mar 9;127(9):1947-1955.
Engberg O, Bocicchio A, Brandner AF, Gupta A, Dey S, Maiti S, Böckmann RA, Huster D. Serotonin alters the phase equilibrium of a ternary mixture of phospholipids and cholesterol. Front Physiol. 2022. 11:578868.
Musabirova G, Engberg O, Gupta A, Roy DS, Maiti S, Huster D. Serotonergic drugs modulate the phase behavior of complex lipid bilayers. Biochimie. 2022; 18:S0300-9084(22)00101-8.
Tang T, Tan Q, Han S, Diemar A, Löbner K, Wang H, Schüß C, Behr V, Mörl K, Wang M, Chu X, Yi C, Keller M, Kofoed J, Reedtz-Runge S, Kaiser A, Beck-Sickinger AG, Zhao Q, Wu B. Receptor-specific recognition of NPY peptides revealed by structures of NPY receptors. Sci Adv. 2022; 18:eabm1232.
Vu O, Bender BJ, Pankewitz L, Huster S, Beck-Sickinger AG, Meiler J. The structural basis of peptide binding at class A G protein-coupled receptors. Molecules. 2022. 27:210.
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.
Tang T, Hartig C, Chen Q, Zhao W, Kaiser A, Zhang X, Zhang H, Qu H, Yi C, Ma L, Han S, Zhao Q, Beck-Sickinger AG, Wu B. Structural basis for ligand recognition of the neuropeptide Y Y2 receptor. Nat Commun. 2021; 12:737.
Sánchez MF, Els-Heindl S, Beck-Sickinger AG, Wienke R, Tampé R. Photoinduced receptor confinement drives ligand-independent GPCR signaling. Science. 2021; 371:eabb7657.
Wanka L, Behr V, Beck-Sickinger AG. Arrestin-dependent internalization of rhodopsin-like G protein-coupled receptors. Biol Chem. 2021; 403:133-149.
Bocicchio A, Brandner AF, Engberg O, Huster D, Böckmann R. Spontaneous membrane nanodomain formation in the absence and in the presence of the neurotransmitter serotonin. Front Cell Dev Biol. 2020. 8:601145.
Huster D. Structural dynamics of G protein-coupled receptors in lipid membranes investigated by solid-state NMR spectroscopy. in Solid-State NMR: Applications in Biomembrane Structure. Editors Frances Separovic and Marc-Antoine Sani IOP Publishing Ltd, 2020 (ISBN: 978-0-7503-2530-1), 11-1-11-27.
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. 2020; 25(18), 4143.
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.
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.
