Structure determination of inactive-state GPCRs with a universal nanobody

Michael J. Robertson, Makaía M. Papasergi-Scott, Feng He, Alpay B. Seven, Justin G. Meyerowitz, Ouliana Panova, Maria Claudia Peroto, Tao Che, Georgios Skiniotis

Research output: Contribution to journalArticlepeer-review

58 Scopus citations

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.

Original languageEnglish
Pages (from-to)1188-1195
Number of pages8
JournalNature Structural and Molecular Biology
Volume29
Issue number12
DOIs
StatePublished - Dec 2022

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