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TitleStructure determination of inactive-state GPCRs with a universal nanobody.
Journal, issue, pagesNat Struct Mol Biol, Vol. 29, Issue 12, Page 1188-1195, Year 2022
Publish dateNov 17, 2022
AuthorsMichael J Robertson / Makaía M Papasergi-Scott / Feng He / Alpay B Seven / Justin G Meyerowitz / Ouliana Panova / Maria Claudia Peroto / Tao Che / Georgios Skiniotis /
PubMed AbstractCryogenic 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 ...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.
External linksNat Struct Mol Biol / PubMed:36396979 / PubMed Central
MethodsEM (single particle)
Resolution2.4 - 3.1 Å
Structure data

26589

7ul2

EMDB-26589, PDB-7ul2:
CryoEM Structure of Inactive NTSR1 Bound to SR48692 and Nb6
Method: EM (single particle) / Resolution: 2.4 Å

26590

7ul3

EMDB-26590, PDB-7ul3:
CryoEM Structure of Inactive H2R Bound to Famotidine, Nb6M, and NabFab
Method: EM (single particle) / Resolution: 3.0 Å

26591

7ul4

EMDB-26591, PDB-7ul4:
CryoEM Structure of Inactive MOR Bound to Alvimopan and Mb6
Method: EM (single particle) / Resolution: 2.8 Å

26592

7ul5

EMDB-26592, PDB-7ul5:
CryoEM Structure of Inactive SSTR2 bound to Nb6
Method: EM (single particle) / Resolution: 3.1 Å

Chemicals

ChemComp-Q6Q:
2-[[1-(7-chloranylquinolin-4-yl)-5-(2,6-dimethoxyphenyl)pyrazol-3-yl]carbonylamino]adamantane-2-carboxylic acid

ChemComp-NA:
Unknown entry

ChemComp-HOH:
WATER

ChemComp-FO9:
famotidine

ChemComp-NG0:
N-[(2S)-2-{[(3R,4R)-4-(3-hydroxyphenyl)-3,4-dimethylpiperidin-1-yl]methyl}-3-phenylpropanoyl]glycine / antagonist*YM

Source
  • homo sapiens (human)
  • lama glama (llama)
  • synthetic construct (others)
  • mus musculus (house mouse)
KeywordsMEMBRANE PROTEIN / Antagonist / Complex

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