Conformational plasticity of the intracellular cavity of GPCR−G-protein complexes leads to G-protein promiscuity and selectivity

Manbir Sandhu; Anja M. Touma; Matthew Dysthe; Fredrik Sadler; Sivaraj Sivaramakrishnan; Nagarajan Vaidehi

doi:10.1073/pnas.1820944116

Conformational plasticity of the intracellular cavity of GPCR−G-protein complexes leads to G-protein promiscuity and selectivity

Manbir Sandhu, Anja M. Touma, Matthew Dysthe, Fredrik Sadler, Sivaraj Sivaramakrishnan, Nagarajan Vaidehi

Research output: Contribution to journal › Article › peer-review

56 Scopus citations

Abstract

While the dynamics of the intracellular surface in agonist-stimulated GPCRs is well studied, the impact of GPCR dynamics on G-protein selectivity remains unclear. Here, we combine molecular dynamics simulations with live-cell FRET and secondary messenger measurements, for 21 GPCR−G-protein combinations, to advance a dynamic model of the GPCR−G-protein interface. Our data show C terminus peptides of Gα_s, Gαi, and Gα_q proteins assume a small ensemble of unique orientations when coupled to their cognate GPCRs, similar to the variations observed in 3D structures of GPCR−G-protein complexes. The noncognate G proteins interface with latent intracellular GPCR cavities but dissociate due to weak and unstable interactions. Three predicted mutations in β₂-adrenergic receptor stabilize binding of noncognate Gα_q protein in its latent cavity, allowing promiscuous signaling through both Gα_s and Gα_q in a dose-dependent manner. This demonstrates that latent GPCR cavities can be evolved, by design or nature, to tune G-protein selectivity, giving insights to pluridimensional GPCR signaling.

Original language	English (US)
Pages (from-to)	11956-11965
Number of pages	10
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	116
Issue number	24
DOIs	https://doi.org/10.1073/pnas.1820944116
State	Published - Jun 11 2019

Bibliographical note

Funding Information:
Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.).

Funding Information:
ACKNOWLEDGMENTS. Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.).

Publisher Copyright:
© 2019 National Academy of Sciences. All rights reserved.

Keywords

G-protein−coupled receptor
GPCR
dynamics
functional selectivity
structural plasticity

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10.1073/pnas.1820944116

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Conformational plasticity of the intracellular cavity of GPCR−G-protein complexes leads to G-protein promiscuity and selectivity. / Sandhu, Manbir; Touma, Anja M.; Dysthe, Matthew et al.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 24, 11.06.2019, p. 11956-11965.

Research output: Contribution to journal › Article › peer-review

Sandhu, M, Touma, AM, Dysthe, M, Sadler, F, Sivaramakrishnan, S & Vaidehi, N 2019, 'Conformational plasticity of the intracellular cavity of GPCR−G-protein complexes leads to G-protein promiscuity and selectivity', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 24, pp. 11956-11965. https://doi.org/10.1073/pnas.1820944116

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abstract = "While the dynamics of the intracellular surface in agonist-stimulated GPCRs is well studied, the impact of GPCR dynamics on G-protein selectivity remains unclear. Here, we combine molecular dynamics simulations with live-cell FRET and secondary messenger measurements, for 21 GPCR−G-protein combinations, to advance a dynamic model of the GPCR−G-protein interface. Our data show C terminus peptides of Gαs, Gαi, and Gαq proteins assume a small ensemble of unique orientations when coupled to their cognate GPCRs, similar to the variations observed in 3D structures of GPCR−G-protein complexes. The noncognate G proteins interface with latent intracellular GPCR cavities but dissociate due to weak and unstable interactions. Three predicted mutations in β2-adrenergic receptor stabilize binding of noncognate Gαq protein in its latent cavity, allowing promiscuous signaling through both Gαs and Gαq in a dose-dependent manner. This demonstrates that latent GPCR cavities can be evolved, by design or nature, to tune G-protein selectivity, giving insights to pluridimensional GPCR signaling.",

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note = "Funding Information: Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.). Funding Information: ACKNOWLEDGMENTS. Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.). Publisher Copyright: {\textcopyright} 2019 National Academy of Sciences. All rights reserved.",

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AU - Sadler, Fredrik

AU - Sivaramakrishnan, Sivaraj

AU - Vaidehi, Nagarajan

N1 - Funding Information: Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.). Funding Information: ACKNOWLEDGMENTS. Research in this publication is supported by Grant NIH-R01GM117923 (to N.V. and S.S.) and Maximizing Investigator Research Award Grant NIH-R35GM126940 (to S.S.). Publisher Copyright: © 2019 National Academy of Sciences. All rights reserved.

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N2 - While the dynamics of the intracellular surface in agonist-stimulated GPCRs is well studied, the impact of GPCR dynamics on G-protein selectivity remains unclear. Here, we combine molecular dynamics simulations with live-cell FRET and secondary messenger measurements, for 21 GPCR−G-protein combinations, to advance a dynamic model of the GPCR−G-protein interface. Our data show C terminus peptides of Gαs, Gαi, and Gαq proteins assume a small ensemble of unique orientations when coupled to their cognate GPCRs, similar to the variations observed in 3D structures of GPCR−G-protein complexes. The noncognate G proteins interface with latent intracellular GPCR cavities but dissociate due to weak and unstable interactions. Three predicted mutations in β2-adrenergic receptor stabilize binding of noncognate Gαq protein in its latent cavity, allowing promiscuous signaling through both Gαs and Gαq in a dose-dependent manner. This demonstrates that latent GPCR cavities can be evolved, by design or nature, to tune G-protein selectivity, giving insights to pluridimensional GPCR signaling.

AB - While the dynamics of the intracellular surface in agonist-stimulated GPCRs is well studied, the impact of GPCR dynamics on G-protein selectivity remains unclear. Here, we combine molecular dynamics simulations with live-cell FRET and secondary messenger measurements, for 21 GPCR−G-protein combinations, to advance a dynamic model of the GPCR−G-protein interface. Our data show C terminus peptides of Gαs, Gαi, and Gαq proteins assume a small ensemble of unique orientations when coupled to their cognate GPCRs, similar to the variations observed in 3D structures of GPCR−G-protein complexes. The noncognate G proteins interface with latent intracellular GPCR cavities but dissociate due to weak and unstable interactions. Three predicted mutations in β2-adrenergic receptor stabilize binding of noncognate Gαq protein in its latent cavity, allowing promiscuous signaling through both Gαs and Gαq in a dose-dependent manner. This demonstrates that latent GPCR cavities can be evolved, by design or nature, to tune G-protein selectivity, giving insights to pluridimensional GPCR signaling.

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Conformational plasticity of the intracellular cavity of GPCR−G-protein complexes leads to G-protein promiscuity and selectivity

Abstract

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