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. Author manuscript; available in PMC: 2026 Jan 1.
Published in final edited form as: Trends Biochem Sci. 2024 Nov 9;50(1):84–85. doi: 10.1016/j.tibs.2024.10.007

NMR spectroscopy reveals insights into mechanisms of GPCR signaling

Larissa O Silva 1,#, Anuradha V Wijesekara 1,#, Matthew T Eddy 1,*
PMCID: PMC11698642  NIHMSID: NIHMS2031108  PMID: 39523165

Advantages

Native proteins: Native or nearly-native protein sequences are more often utilized

Non-perturbative: NMR probes are stable isotopes with minimal to no perturbation of the protein structure

Molecular sizes: There is no inherent lower or upper limit to the molecular size of proteins if considering both solution and solid-state NMR capabilities

Diverse environments: data can be measured in aqueous environments via solution NMR or in solids via solid-state NMR across many membrane or membrane-mimetic environments

Physiological temperatures: samples can be measured across range of temperatures, including human body temperature

Quantitative measurements: kinetics of exchange processes can be quantitatively measured over many orders of magnitude in time

Challenges

Sensitivity and sample purification: NMR experiments often require 1 milligram or more of purified protein

Stable isotope incorporation: protein-observed experiments usually require incorporation of one or more stable-isotopes

Instrumentation requirements: solid-state and solution NMR experiments typically require different radiofrequency probes and spectrometer configurations

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Nuclear magnetic resonance (NMR) spectroscopy is a powerful technique used to study challenging membrane proteins, including G protein-coupled receptors (GPCRs). NMR provides detailed insights into a wide range of receptor properties, including ligand-stimulated signaling mechanisms and pathways, receptor-lipid and receptor-ligand interactions, and quantitative measurements of conformational exchange rates and receptor structure.

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Two complementary approaches for incorporating NMR-visible stable-isotopes are chemical modification and isotope enrichment through cell culture media. The labeled receptors are subsequently purified and reconstituted into membrane-mimetic systems, such as detergents or lipid nanodiscs, or into larger lipid vesicles that more closely resemble native cellular membranes. While detergent and nanodisc samples are typically suited for solution NMR experiments, receptors embedded in lipid vesicles are analyzed using solid-state NMR techniques.

Acknowledgements

The authors acknowledge funding for this work the National Institutes of Health grant number R35GM138291, NSF CAREER award 2339330 (M.T.E.) and NIH T32 training grant T32GM136583 (L.O.S.).

Footnotes

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Declaration of Interests

No interests are declared

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