Local membrane charge regulates β2 adrenergic receptor coupling to Gi3

Strohman, Michael; Maeda, Shoji; Hilger, Daniel; Masureel, Matthieu; Du, Yang; Kobilka, Brian K.

doi:10.1038/s41467-019-10108-0

Cited by 70 publications

(69 citation statements)

References 73 publications

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“…5 compares the spectra of β 2 AR-G s EMPTY and β 2 AR-G i1 EMPTY complexes. Consistent with the previous observation that the β 2 AR-G i1 complex is less stable than the β 2 AR-G s complex ( 29 ), we observed smaller spectral changes in the β 2 AR-G i1 complex sample. The NMR spectral changes of K227 5.56 , K267 6.29 , K273 6.35 , and R328K 7.55 corresponding to the conformational changes of TM5, 6, and 7 are similar to those obtained from the β 2 AR-G s complex; however, peak 2 of K227 5.56 and K273 6.35 and peak 1 of K267 6.29 are notably weaker in the β 2 AR-G i1 complex compared to the β 2 AR-G s complex.…”

Section: Resultssupporting

confidence: 92%

Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy

Batebi³

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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The β2-adrenergic receptor (β2AR) is a prototypical G protein-coupled receptor (GPCR) that preferentially couples to the stimulatory G protein Gs and stimulates cAMP formation. Functional studies have shown that the β2AR also couples to inhibitory G protein Gi, activation of which inhibits cAMP formation [R. P. Xiao, Sci. STKE 2001, re15 (2001)]. A crystal structure of the β2AR-Gs complex revealed the interaction interface of β2AR-Gs and structural changes upon complex formation [S. G. Rasmussen et al., Nature 477, 549–555 (2011)], yet, the dynamic process of the β2AR signaling through Gs and its preferential coupling to Gs over Gi is still not fully understood. Here, we utilize solution nuclear magnetic resonance (NMR) spectroscopy and supporting molecular dynamics (MD) simulations to monitor the conformational changes in the G protein coupling interface of the β2AR in response to the full agonist BI-167107 and Gs and Gi1. These results show that BI-167107 stabilizes conformational changes in four transmembrane segments (TM4, TM5, TM6, and TM7) prior to coupling to a G protein, and that the agonist-bound receptor conformation is different from the G protein coupled state. While most of the conformational changes observed in the β2AR are qualitatively the same for Gs and Gi1, we detected distinct differences between the β2AR-Gs and the β2AR-Gi1 complex in intracellular loop 2 (ICL2). Interactions with ICL2 are essential for activation of Gs. These differences between the β2AR-Gs and β2AR-Gi1 complexes in ICL2 may be key determinants for G protein coupling selectivity.

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Section: Resultssupporting

confidence: 92%

Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy

Batebi³

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…This includes commonly used neutral detergents, as well as zwitterionic membrane lipids used in lipid nanodiscs or nanoparticles. With the ambition to mimic cellular membranes, it is reasonable to include negatively charged head groups in the membrane mimetic [ 88 , 89 ]. Ψ I comprises the electric potentials of the hydrocarbon core of the lipid membrane.…”

Section: Interactions Of Gpcrs With Their Membrane Environmentmentioning

confidence: 99%

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

et al. 2020

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Over the past decade, the vast amount of information generated through structural and biophysical studies of GPCRs has provided unprecedented mechanistic insight into the complex signalling behaviour of these receptors. With this recent information surge, it has also become increasingly apparent that in order to reproduce the various effects that lipids and membranes exert on the biological function for these allosteric receptors, in vitro studies of GPCRs need to be conducted under conditions that adequately approximate the native lipid bilayer environment. In the first part of this review, we assess some of the more general effects that a membrane environment exerts on lipid bilayer-embedded proteins such as GPCRs. This is then followed by the consideration of more specific effects, including stoichiometric interactions with specific lipid subtypes. In the final section, we survey a range of different membrane mimetics that are currently used for in vitro studies, with a focus on NMR applications.

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“…PE, by contrast, favours an inactive receptor state. In a subsequent study, it was found that nanodiscs ApoA-1 POPC/POPG (3:2) and BPL/POPC/POPG (1.07:1.5:1) Ligand binding and G protein coupling of monomeric receptor (Kuszak et al 2009) Adenosine A 2A -receptor (A containing PG and PS headgroups modulate β 2 AR activation by the G s heterotrimer but surprisingly are inhibitory to β 2 AR activation by the G i3 heterotrimer in the absence of divalent cations (Strohman et al 2019). Instead, PE facilitates β 2 AR activation by G i3 , demonstrating that one type of lipid can exert different effects at the levels of GPCR conformation and GPCR-G protein coupling.…”

Section: Nanodiscsmentioning

confidence: 99%

Lipid nanoparticle technologies for the study of G protein-coupled receptors in lipid environments

Lavington

Watts

2020

Biophys Rev

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G protein-coupled receptors (GPCRs) are a large family of integral membrane proteins which conduct a wide range of biological roles and represent significant drug targets. Most biophysical and structural studies of GPCRs have been conducted on detergent-solubilised receptors, and it is clear that detergents can have detrimental effects on GPCR function. Simultaneously, there is increasing appreciation of roles for specific lipids in modulation of GPCR function. Lipid nanoparticles such as nanodiscs and styrene maleic acid lipid particles (SMALPs) offer opportunities to study integral membrane proteins in lipid environments, in a form that is soluble and amenable to structural and biophysical experiments. Here, we review the application of lipid nanoparticle technologies to the study of GPCRs, assessing the relative merits and limitations of each system. We highlight how these technologies can provide superior platforms to detergents for structural and biophysical studies of GPCRs and inform on roles for protein-lipid interactions in GPCR function.

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Local membrane charge regulates β2 adrenergic receptor coupling to Gi3

Cited by 70 publications

References 73 publications

Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy

Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

Lipid nanoparticle technologies for the study of G protein-coupled receptors in lipid environments

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Local membrane charge regulates β2 adrenergic receptor coupling to Gi3

Cited by 70 publications

References 73 publications

Analysis of β 2 AR-G s and β 2 AR-G i complex formation by NMR spectroscopy

Analysis of β 2 AR-G s and β 2 AR-G i complex formation by NMR spectroscopy

Structure and Dynamics of GPCRs in Lipid Membranes: Physical Principles and Experimental Approaches

Lipid nanoparticle technologies for the study of G protein-coupled receptors in lipid environments

Contact Info

Product

Resources

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Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy

Analysis of β ₂ AR-G _s and β ₂ AR-G _i complex formation by NMR spectroscopy