A Comprehensive Mutagenesis Screen of the Adhesion GPCR Latrophilin-1/ADGRL1
Olha Nazarko,
Amanuel Kibrom,
Jana Winkler,
Katherine Leon,
Hannah Stoveken,
Gabriel Salzman,
Katarzyna Merdas,
Yue Lu,
Pradnya Narkhede,
Gregory Tall,
Simone Prömel,
Demet Araç
Affiliations
Olha Nazarko
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Amanuel Kibrom
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Jana Winkler
Rudolf Schönheimer Institute of Biochemistry, Molecular Biochemistry, Medical Faculty, University of Leipzig, 04103 Leipzig, Germany
Katherine Leon
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Hannah Stoveken
Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA
Gabriel Salzman
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Katarzyna Merdas
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Yue Lu
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Pradnya Narkhede
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA
Gregory Tall
Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA
Simone Prömel
Rudolf Schönheimer Institute of Biochemistry, Molecular Biochemistry, Medical Faculty, University of Leipzig, 04103 Leipzig, Germany
Demet Araç
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637, USA; Grossman Institute for Neuroscience, Quantitative Biology and Human Behavior, The University of Chicago, Chicago, IL 60637, USA; Corresponding author
Summary: Adhesion G-protein-coupled receptors (aGPCRs) play critical roles in diverse cellular processes in neurobiology, development, immunity, and numerous diseases. The lack of molecular understanding of their activation mechanisms, especially with regard to the transmembrane domains, hampers further studies to facilitate aGPCR-targeted drug development. Latrophilin-1/ADGRL1 is a model aGPCR that regulates synapse formation and embryogenesis, and its mutations are associated with cancer and attention-deficit/hyperactivity disorder. Here, we established functional assays to monitor latrophilin-1 function and showed the activation of latrophilin-1 by its endogenous agonist peptide. Via a comprehensive mutagenesis screen, we identified transmembrane domain residues essential for latrophilin-1 basal activity and for agonist peptide response. Strikingly, a cancer-associated mutation exhibited increased basal activity and failed to rescue the embryonic developmental phenotype in transgenic worms. These results provide a mechanistic foundation for future aGPCR-targeted drug design. : Molecular Biology; Membrane Architecture; Protein Structure Aspects Subject Areas: Molecular Biology, Membrane Architecture, Protein Structure Aspects
