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Isoform- and ligand-specific modulation of the adhesion GPCR ADGRL3/Latrophilin3 by a synthetic binder

Kordon, Szymon P. and Dutka, Przemysław and Adamska, Justyna M. and Bandekar, Sumit J. and Leon, Katherine and Adams, Brock and Li, Jingxian and Kossiakoff, Anthony A. and Araç, Demet (2022) Isoform- and ligand-specific modulation of the adhesion GPCR ADGRL3/Latrophilin3 by a synthetic binder. . (Unpublished) https://resolver.caltech.edu/CaltechAUTHORS:20220722-769606000

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Abstract

Adhesion G protein-coupled receptors (aGPCRs) are cell-surface proteins with large extracellular regions that bind to multiple ligands to regulate key biological functions including neurodevelopment and organogenesis. Modulating a single function of a specific aGPCR isoform while affecting no other function and no other receptor is not trivial. Here, we engineered an antibody, termed LK30, that binds to the extracellular region of the aGPCR ADGRL3, and specifically acts as an agonist for ADGRL3 but not for its isoform, ADGRL1. The LK30/ADGRL3 complex structure revealed that the LK30 binding site on ADGRL3 overlaps with the binding site for an ADGRL3 ligand – teneurin. In cellular-adhesion assays, LK30 specifically broke the trans-cellular interaction of ADGRL3 with teneurin, but not with another ADGRL3 ligand – FLRT3. Our work provides proof of concept for the modulation of isoform- and ligand-specific aGPCR functions using unique tools, and thus establishes a foundation for the development of fine-tuned aGPCR-targeted therapeutics.


Item Type:Report or Paper (Discussion Paper)
Related URLs:
URLURL TypeDescription
https://doi.org/10.1101/2022.07.20.500857DOIArticle
ORCID:
AuthorORCID
Kordon, Szymon P.0000-0002-3770-0743
Dutka, Przemysław0000-0003-3819-1618
Bandekar, Sumit J.0000-0003-2728-7366
Leon, Katherine0000-0002-0870-5000
Kossiakoff, Anthony A.0000-0003-3174-9359
Araç, Demet0000-0002-5444-5799
Additional Information:The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. Version 1 - July 21, 2022; Version 2 - July 22, 2022. We thank Engin Özkan for the use of flow cytometer and all members of the Araç lab for helpful discussions. We also thank the staff at the Advanced Photon Source (APS) at Argonne National Labs (ANL). GM/CA@APS has been funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006, P30GM138396). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The Eiger 16M detector at GM/CA-XSD was funded by NIH grant S10 OD012289. This work was supported by grants R01 GM120322 (to D.A.), R01 GM134035-01 (to D.A.) and GM117372 (to A.A.K.). Author Contributions: S.P.K. and D.A. designed all experiments and interpreted results. S.P.K. expressed and purified all proteins (with assistance from B.A. and K.L.). P.D. carried out phage display selection and sABs characterization. S.P.K. and J.M.A. performed flow cytometry experiments and cell-based signaling assays. and S.P.K. performed crystallography experiments (with assistance from B.A.) and structure determination. J.L. participated in cell-cell aggregation experiments. S.P.K. and D.A. wrote the manuscript with assistance from A.A.K., S.J.B. and P.D.. D.A. and A.A.K. supervised the project. The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
NIHACB-12002
NIHAGM-12006
NIHP30GM138396
Department of Energy (DOE)DE-AC02-06CH11357
NIHS10 OD012289
NIHR01 GM120322
NIHR01 GM134035-01
NIHGM117372
Subject Keywords:adhesion GPCR, latrophilin, teneurin, synapse, protein engineering
DOI:10.1101/2022.07.20.500857
Record Number:CaltechAUTHORS:20220722-769606000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220722-769606000
Official Citation:Isoform- and ligand-specific modulation of the adhesion GPCR ADGRL3/Latrophilin3 by a synthetic binder. Szymon P. Kordon, Przemysław Dutka, Justyna M. Adamska, Sumit J. Bandekar, Katherine Leon, Brock Adams, Jingxian Li, Anthony A. Kossiakoff, Demet Araç. bioRxiv 2022.07.20.500857; doi: https://doi.org/10.1101/2022.07.20.500857
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:115802
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:26 Jul 2022 17:30
Last Modified:02 Aug 2022 19:59

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