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Structure determination of inactive-state GPCRs with a universal nanobody.

Robertson MJ, Papasergi-Scott MM, He F, Seven AB, Meyerowitz JG, Panova O, Peroto MC, Che T, Skiniotis G
Nat Struct Mol Biol (2022)
Related entries: 7ul3, 7ul4, 7ul5

Cited: 42 times
EuropePMC logo PMID: 36396979

Abstract

Cryogenic electron microscopy (cryo-EM) has widened the field of structure-based drug discovery by allowing for routine determination of membrane protein structures previously intractable. Despite representing one of the largest classes of therapeutic targets, most inactive-state G protein-coupled receptors (GPCRs) have remained inaccessible for cryo-EM because their small size and membrane-embedded nature impedes projection alignment for high-resolution map reconstructions. Here we demonstrate that the same single-chain camelid antibody (nanobody) recognizing a grafted intracellular loop can be used to obtain cryo-EM structures of inactive-state GPCRs at resolutions comparable or better than those obtained by X-ray crystallography. Using this approach, we obtained structures of neurotensin 1 receptor bound to antagonist SR48692, μ-opioid receptor bound to alvimopan, apo somatostatin receptor 2 and histamine receptor 2 bound to famotidine. We expect this rapid, straightforward approach to facilitate the broad exploration of GPCR inactive states without the need for extensive engineering and crystallization.

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  17. Mechanistic Elucidation of Activation/Deactivation Signal Transduction within Neurotensin Receptor 1 Triggered by 'Driver Chemical Groups' of Modulators: A Comparative Molecular Dynamics Simulation. Lu X, Shi X, Fan J, Li M, Zhang Y, Lu S, Xu G, Chen Z. Pharmaceutics 15 2000 (2023)
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  22. Insight into the Mode of Action of 8-Hydroxyquinoline-Based Blockers on the Histamine Receptor 2. Patel A, Marquez-Gomez PL, Torp LR, Gao L, Peralta-Yahya P. Biosensors (Basel) 13 571 (2023)
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