Avaliações PREreview 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

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

Publicado: 27 de julho de 2023
Servidor: bioRxiv
DOI: 10.1101/2023.07.27.550782

Resumo

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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Avaliação PREreview feita por Emelyne M. Pacull et al. do/a Biophysics Leipzig University

De autoria de Emelyne M. Pacull, Jeannette Laugwitz e Matthias Voitel do/a 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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