PREreviews de Cryo-EM structure of cell-free synthesized human histamine H 2 receptor coupled to heterotrimeric G s protein in lipid nanodisc environment

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Cryo-EM structure of cell-free synthesized human histamine H ₂ receptor coupled to heterotrimeric G _s protein in lipid nanodisc environment

par Zoe Köck, Kilian Schnelle, Margherita Persechino, Simon Umbach, Hannes Schihada, Dovile Januliene, Kristian Parey, Steffen Pockes, Peter Kolb, Volker Dötsch, Arne Möller, Daniel Hilger et Frank Bernhard

Publié: 27 juillet 2023
Serveur de preprints: bioRxiv
DOI: 10.1101/2023.07.27.550782

Résumé

Here we describe the cryo-electron microscopy structure of the human histamine 2 receptor (H ₂ R) in an active conformation with bound histamine and in complex with G _s heterotrimeric protein at an overall resolution of 3.4 Å. The complex was generated by cotranslational insertion into preformed nanodisc membranes using cell-free synthesis in E. coli lysates. It is the first structure obtained by this detergent-free strategy and the first GPCR/G _s complex structure in lipid environment. Structural comparison with the inactive conformation of H ₂ R and the inactive and G _q -coupled active state of H ₁ R together with structure-guided functional experiments reveal molecular insights into the specificity of ligand binding and G protein coupling for this receptor family. We demonstrate lipid-modulated folding of cell-free synthesized H ₂ R, its agonist-dependent internalization and its interaction with endogenously synthesized H ₁ R and H ₂ R in HEK293 cells by applying a recently developed nanotransfer technique.

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PREreview Emelyne M. Pacull et al. du club Biophysics Leipzig University

Rédigé par Emelyne M. Pacull, Jeannette Laugwitz et Matthias Voitel du Biophysics Leipzig University

Summary of the paper:

In this paper, the authors report on the expression of the human histamine 2 receptor (H₂R) using the cell-free (CF) expression procedure with co-translational insertion into nanodiscs and formation of a complex with the endogenous ligand histamine, the Gs protein and the Gs…

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