Project B06 – Dynamic modulation of adhesion GPCR function through complex formation
Adhesion GPCR (aGPCR) use unusual molecular strategies to modulate the physiological functions of the expressing and neighboring cells. Our recent work demonstrated that aGPCR isoforms form homo and heteromeric assemblies to tune cellular mechano-responsiveness. Building on this finding, we now aim to identify the molecular interfaces that enable complex formation and explore the molecular mechanisms that underlie their activity and signaling facility. In this context, we will investigate if and how the newly identified ligand TLR 8 (toll-like receptor 8, Tollo) is involved and how cell autonomy of this aGPCR-ligand pair is organized. Moreover, we intend to decipher the function of additional aGPCR-ligand pairs in vivo in their native cellular context. For these purposes, we will generate a FRET-based toolkit of intra and intermolecular aGPCR and non-GPCR sensors, use established signaling assays and proteinbiochemical protocols in combination with a series of in vivo readouts including Ca2+ imaging and electrophysiology.
The activity profile and signaling capacity of adhesion GPCRs (aGPCR) may be modulated through crosstalk between different cell surface molecules and may be triggered and/or modulated by mechanical input. We will identify ligand-receptor complexes of aGPCRs through in vitro and in vivo orthogonal labeling and photo-crosslinking as well as intermolecular FRET approaches, respectively. In a further tier of experiments, we will investigate if putative interactors affect aGPCR signaling in a mechano-dependent fashion, test if mechanical stimuli serve as a switch for aGPCR dimerization/oligomerization and investigate how the viscoelastic properties of these interactions correlate with aGPCR activity.
Contact
Prof. Dr. D.Phil. (Oxon) Tobias Langenhan (Project Leader)
Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig
Phone +49 341 97 22101
E-Mail
Web langenhan-lab.org/
Dr. Nicole Scholz (Project Leader)
Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig
Phone +49 341 97 22145
E-Mail
Web uniklinikum-leipzig.de/ag-scholz
Anne-Kristin Dahse (PhD Student)
Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig
Anne Bormann (PhD Student)
Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig
Michal Mitro (PhD Student)
Leipzig University, Faculty of Medicine
Rudolf Schönheimer Institute of Biochemistry
Johannisallee 30, D-04103 Leipzig
Resources
- in vitro and in vivo orthogonal labeling and photo-crosslinking as well as intermolecular FRET approaches
- Drosophila melanogaster as an in vivo model
- Proteinbiochemical approaches as well as behavioral and functional readouts
Publications
Buhlan M, Ljaschenko D, Scholz N#, Langenhan T#. Experimental modulation of physiological force application on leg joint neurons in intact Drosophila melanogaster. Nature Protocols. 2024; 19:113-126. DOI: 10.1038/s41596-023-00907-7.
Vieira Contreras F, Auger GM, Müller L, Richter V, Huetteroth W, Seufert F, Hildebrand PW, Scholz N, Thum AS, Ljaschenko D, Blanco-Redondo B, Langenhan T. The adhesion G-protein-coupled receptor mayo/CG11318 controls midgut development in Drosophila. Cell Rep. 2024 Jan 4;43(1):113640. doi: 10.1016/j.celrep.2023.113640.
Fu C, Huang W, Tang Q, Niu M, Guo S, Langenhan T, Song G, Yan J. Unveiling Mechanical Activation: GAIN Domain Unfolding and Dissociation in Adhesion GPCRs. Nano Lett. 2023; 25;23(20):9179-9186.
Mrestani A, Dannhäuser S, Pauli M, Kollmannsberger P, Hübsch M, Morris L, Langenhan T, Heckmann M, Paul MM. Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging. Life Sci Alliance. 2023 Sep 11;6(12):e202302021. doi: 10.26508/lsa.202302021. PMID: 37696575; PMCID: PMC10494931.
Scholz N#, Dahse AK, Kemkemer M, Bormann A, Auger GM, Vieira Contreras F, Ernst LF, Staake H, Körner MB, Buhlan M, Meyer-Mölck A, Chung YK, Blanco-Redondo B, Klose F, Jarboui MA, Ljaschenko D, Bigl M, Langenhan T#. Molecular sensing of mechano- and ligand-dependent adhesion GPCR dissociation. Nature. 2023; 615:945-953.
Strutt H, Warrington S, Madathil ACK, Langenhan T, Strutt D Molecular symmetry breaking in the Frizzled-dependent planar polarity pathway. Current Biology. 2023. Curr Biol 33:5340-5354.e6.
Thane M, Paisios E, Stöter T, Krüger AR, Gläß S, Dahse AK, Scholz N, Gerber B, Lehmann DJ, Schleyer M. High-resolution analysis of individual Drosophila melanogaster larvae uncovers individual variability in locomotion and its neurogenetic modulation. Open Biol. 2023 Apr;13(4):220308.
Götze KJ, Mrestani A, Beckmann P, Krohn K, Le Duc D, Velluva A, Böhme MA, Heckmann M, Abou Jamra R, Lemke JR, Bläker H, Scholz N, Ljaschenko D, Langenhan T. Improving one-step scarless genome editing in Drosophila melanogaster by combining ovoD co-CRISPR selection with sgRNA target site masking. Biol Methods Protoc. 2022 Jan 12;7(1):bpac003. doi: 10.1093/biomethods/bpac003 . PMID: 35087953 ; PMCID: PMC8789338.
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. Mol Cell. 2021 Mar 4;81(5):905-921.e5. doi: 10.1016/j.molcel.2020.12.042. Epub 2021 Jan 25.
Smolič T, Tavčar P, Horvat A, Černe U, Halužan Vasle A, Tratnjek L, Kreft ME, Scholz N, Matis M, Petan T, Zorec R, Vardjan N. Astrocytes in stress accumulate lipid droplets. Glia. 2021 Feb 20. doi: 10.1002/glia.23978. Epub ahead of print. PMID: 33609060. https://doi.org/10.1002/glia.23978.
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]
Scholz N*, Ehmann N*, Sachidanandan D, Imig C, Cooper BH, Jahn O, Reim K, Brose N, Meyer J, Lamberty M, Altrichter S, Bormann A, Hallermann S, Pauli M, Heckmann M, Stigloher C, Langenhan T#, Kittel RJ#. Complexin cooperates with Bruchpilot to tether synaptic vesicles to the active zone cytomatrix. J Cell Biol. 2019; 218:1011-26.
Scholz N, Langenhan T, Schöneberg T. Revisiting the classification of adhesion GPCRs. Ann N Y Acad Sci. 2019; 1456: 80-95.
Blanco Redondo B, Langenhan T#. Parallel Genomic Engineering of Two Drosophila Genes Using Orthogonal attB/attP Sites. G3 (Bethesda). 2018; 8:3109–3118.
Scholz N. Cancer Cell Mechanics: Adhesion G Protein-coupled Receptors in Action? Front Oncol. 2018; 8:59.
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. 2017; 6:e28360.
Langenhan T#, Piao X#, Monk KR#. Adhesion G protein-coupled receptors in nervous system development and disease. Nat Rev Neurosci. 2016; 17:550-61.
Scholz N#, Monk KR, Kittel RJ, Langenhan T#. Adhesion GPCRs as a Putative Class of Metabotropic Mechanosensors. Handb Exp Pharmacol. 2016; 234:221-47.
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.
Langenhan T*, Aust G*, Hamann J*. Sticky Signaling-Adhesion Class G Protein-Coupled Receptors Take the Stage. Sci Signal. 2013; 6:re3.
Prömel S#, Waller-Evans H, Dixon J, Zahn D, Colledge WH, Doran J, Carlton MBL, Grosse J, Schöneberg T, Russ AP, Langenhan T#. Characterization and functional study of a cluster of four highly conserved orphan adhesion-GPCR in mouse. Dev Dyn. 2012b; 241:1591-602.
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; 2:321-31.
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.
