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Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors
Article

Specific Engineered G Protein Coupling to Histamine Receptors Revealed from Cellular Assay Experiments and Accelerated Molecular Dynamics Simulations

1
Institute of Pharmacy, Faculty of Chemistry and Pharmacy, University of Regensburg, 93040 Regensburg, Germany
2
Bioinformatik, Institut für Biochemie, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Fahrstraße 17, 91054 Erlangen, Germany
3
Department of Computational Biology and Molecular Biosciences, University of Kansas, Lawrence, KS 66047, USA
4
Erlangen National High Performance Computing Center ([email protected]), Friedrich-Alexander-University Erlangen-Nürnberg (FAU), 91058 Erlangen, Germany
*
Authors to whom correspondence should be addressed.
Academic Editor: Paul Chazot
Int. J. Mol. Sci. 2021, 22(18), 10047; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms221810047
Received: 12 August 2021 / Revised: 15 September 2021 / Accepted: 15 September 2021 / Published: 17 September 2021
(This article belongs to the Special Issue Molecular Biology of Histamine Systems)
G protein-coupled receptors (GPCRs) are targets of extracellular stimuli and hence occupy a key position in drug discovery. By specific and not yet fully elucidated coupling profiles with α subunits of distinct G protein families, they regulate cellular responses. The histamine H2 and H4 receptors (H2R and H4R) are prominent members of Gs- and Gi-coupled GPCRs. Nevertheless, promiscuous G protein and selective Gi signaling have been reported for the H2R and H4R, respectively, the molecular mechanism of which remained unclear. Using a combination of cellular experimental assays and Gaussian accelerated molecular dynamics (GaMD) simulations, we investigated the coupling profiles of the H2R and H4R to engineered mini-G proteins (mG). We obtained coupling profiles of the mGs, mGsi, or mGsq proteins to the H2R and H4R from the mini-G protein recruitment assays using HEK293T cells. Compared to H2R–mGs expressing cells, histamine responses were weaker (pEC50, Emax) for H2R–mGsi and –mGsq. By contrast, the H4R selectively bound to mGsi. Similarly, in all-atom GaMD simulations, we observed a preferential binding of H2R to mGs and H4R to mGsi revealed by the structural flexibility and free energy landscapes of the complexes. Although the mG α5 helices were consistently located within the HR binding cavity, alternative binding orientations were detected in the complexes. Due to the specific residue interactions, all mG α5 helices of the H2R complexes adopted the Gs-like orientation toward the receptor transmembrane (TM) 6 domain, whereas in H4R complexes, only mGsi was in the Gi-like orientation toward TM2, which was in agreement with Gs- and Gi-coupled GPCRs structures resolved by X-ray/cryo-EM. These cellular and molecular insights support (patho)physiological profiles of the histamine receptors, especially the hitherto little studied H2R function in the brain, as well as of the pharmacological potential of H4R selective drugs. View Full-Text
Keywords: GPCR–G protein coupling profiles; Gaussian accelerated molecular dynamics (GaMD); split-luciferase complementation assay; histamine signaling; histamine H2 receptor; histamine H4 receptor; engineered G proteins GPCR–G protein coupling profiles; Gaussian accelerated molecular dynamics (GaMD); split-luciferase complementation assay; histamine signaling; histamine H2 receptor; histamine H4 receptor; engineered G proteins
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MDPI and ACS Style

Höring, C.; Conrad, M.; Söldner, C.A.; Wang, J.; Sticht, H.; Strasser, A.; Miao, Y. Specific Engineered G Protein Coupling to Histamine Receptors Revealed from Cellular Assay Experiments and Accelerated Molecular Dynamics Simulations. Int. J. Mol. Sci. 2021, 22, 10047. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms221810047

AMA Style

Höring C, Conrad M, Söldner CA, Wang J, Sticht H, Strasser A, Miao Y. Specific Engineered G Protein Coupling to Histamine Receptors Revealed from Cellular Assay Experiments and Accelerated Molecular Dynamics Simulations. International Journal of Molecular Sciences. 2021; 22(18):10047. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms221810047

Chicago/Turabian Style

Höring, Carina, Marcus Conrad, Christian A. Söldner, Jinan Wang, Heinrich Sticht, Andrea Strasser, and Yinglong Miao. 2021. "Specific Engineered G Protein Coupling to Histamine Receptors Revealed from Cellular Assay Experiments and Accelerated Molecular Dynamics Simulations" International Journal of Molecular Sciences 22, no. 18: 10047. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms221810047

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