Project A06 – Enzymology of autoproteolysis and signaling function of the GAIN domain in adhesion GPCRs

Signal transduction via the GAIN (GPCR autoproteolysis inducing) domain is a unique property of adhesion G protein-coupled receptors (aGPCRs). We will systematically investigate the role of active site residues of the GAIN autoprotease on GPCR proteolysis site (GPS) cleavage by expression of the isolated domain, entire ectodomain and full-length aGPCR. These experiments will characterize the influence and role of active site residues, and the complex formed with other extracellular folds and the 7TM part on GAIN autoproteolytic activity. We will determine crystal structures of ectodomains in the pre-cleavage and post-cleavage states. Furthermore, we will investigate how the structure and dynamics of the GAIN domain influences the activation of aGPCRs by release of the Stachel peptide acting as a tethered agonist.

Contact

Prof. Dr. Norbert Sträter (Project Leader)

Leipzig University, Faculty of Chemistry and Mineralogy
Institute for Bioanalytical Chemistry
Deutscher Platz 5, D-04103 Leipzig

Phone +49 341 97 31311
E-Mail

Prof. Dr. Tobias Langenhan, D.Phil. (Oxon) (Project Leader)

Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig

Phone +49 341 97 22101
E-Mail

Resources

  1. crystal structures of ectodomains in the pre-cleavage and post-cleavage states
  2. investigate how the structure and dynamics of the GAIN domain influences the activation of aGPCRs 
  • Microsys 4000-XL crystallization robot
  • Crystal Rock Imager 1000 and Rock Imager FRAP Formulatrix
  • Imaging microscope system FRET
  • Tecan Free Flow Electrophoresis

Publications

Beliu G, Altrichter S, Guixà-González R, Hemberger M, Brauer I, Dahse AK, Scholz N, Wieduwild R, Kuhlemann A, Batebi H, Seufert F, Pérez-Hernández G, Hildebrand PW, Sauer M, Langenhan T. Tethered agonist exposure in intact adhesion/class B2 GPCRs through intrinsic structural flexibility of the GAIN domain. Molecular Cell, 2021. [in press] https://doi.org/10.1016/j.molcel.2020.12.042.

Dannhäuser S, Lux TJ, Hu C, Selcho M, Chen JT, Ehmann N, Sachidanandan D, Stopp S, Pauls D, Pawlak M, Langenhan T, Soba P, Rittner HL, Kittel RJ. Antinociceptive modulation by the adhesion GPCR CIRL promotes mechanosensory signal discrimination. Elife. 2020 Sep 30;9:e56738. doi: 10.7554/eLife.56738 PMID: 32996461; PMCID: PMC7546736.

Mathiasen S, Palmisano T, Perry NA, Stoveken HM, Vizurraga A, McEwen DP, Okashah N, Langenhan T, Inoue A, Lambert NA, Tall GG, Javitch JA. G12/13 is activated by acute tethered agonist exposure in the adhesion GPCR ADGRL3. Nat Chem Biol. 2020 Aug 10. doi: 10.1038/s41589-020-0617-7. Epub ahead of print. Erratum in: Nat Chem Biol. 2020 Aug 17;: PMID: 32778842. [publisher correction: doi: 10.1038/s41589-020-0649-z]

Hamann J, Aust G, Araç D, Engel FB, Formstone C, Fredriksson R, Hall RA, Harty BL, Kirchhoff C, Knapp B, Krishnan A, Liebscher I, Lin HH, Martinelli DC, Monk KR, Peeters MC, Piao X, Prömel S, Schöneberg T, Schwartz TW, Singer K, Stacey M, Ushkaryov YA, Vallon M, Wolfrum U, Wright MW, Xu L, Langenhan T, Schiöth HB. International Union of Basic and Clinical Pharmacology. XCIV. Adhesion G protein-coupled receptors. Pharmacol Rev. 2015; 67:338-67.

Langenhan T, Prömel S, Mestek L, Esmaeili B, Waller-Evans H, Hennig C, Kohara Y, Avery L, Vakonakis I, Schnabel R, Russ AP. Latrophilin signaling links anterior-posterior tissue polarity and oriented cell divisions in the C. elegans embryo. Dev Cell. 2009; 17:494-504.

Arac D, Sträter N, Seiradake E. Understanding the Structural Basis of Adhesion GPCR Functions. Handb Exp Pharmacol. 2016; 234:67-82.

Zebisch M, Krauss M, Schäfer P, Lauble P, Sträter N. Crystallographic snapshots along the reaction pathway of nucleoside triphosphate diphosphohydrolases. Structure. 2013; 21:1460-75.

Langenhan T, Piao X, Monk KR. Adhesion G protein-coupled receptors in nervous system development and disease. Nat Rev Neurosci. 2016; 17:550-61.

Nieberler M, Kittel RJ, Petrenko A, Lin, HH#, Langenhan T#. Control of Adhesion GPCR Function Through Proteolytic Processing. Handb Exp Pharmacol. 2016; 234:83-109.

Knapp K, Zebisch M, Pippel J, El-Tayeb A, Müller CE, Sträter N. Crystal structure of the human ecto-5′-nucleotidase (CD73): insights into the regulation of purinergic signaling. Structure. 2012; 20:2161-73.

Scholz N, Gehring J, Guan C, Ljaschenko D, Fischer R, Lakshmanan V, Kittel RJ#, Langenhan T#. The Adhesion GPCR Latrophilin/CIRL Shapes Mechanosensation. Cell Rep. 2015; 11:866-74.

Scholz N, Guan C, Nieberler M, Grotemeyer A, Maiellaro I, Gao S, Beck S, Pawlak M, Sauer M, Asan E, Rothemund S, Winkler J, Prömel S, Nagel G, Langenhan T, Kittel RJ. Mechano-dependent signaling by Latrophilin/CIRL quenches cAMP in proprioceptive neurons. Elife. 2017; 6:e28360.

Prömel S, Frickenhaus M, Hughes S, Mestek L, Staunton D, Woollard A, Vakonakis I, Schöneberg T, Schnabel R, Russ AP, Langenhan T. The GPS motif is a molecular switch for bimodal activities of adhesion class G protein-coupled receptors. Cell Rep. 2012; 2:321-331.