Molecular mechanism of Gαi activation by non-GPCR proteins with a Gα-Binding and Activating motif

Nature Communications
Alain Ibáñez de OpakuaMikel Garcia-Marcos

Abstract

Heterotrimeric G proteins are quintessential signalling switches activated by nucleotide exchange on Gα. Although activation is predominantly carried out by G-protein-coupled receptors (GPCRs), non-receptor guanine-nucleotide exchange factors (GEFs) have emerged as critical signalling molecules and therapeutic targets. Here we characterize the molecular mechanism of G-protein activation by a family of non-receptor GEFs containing a Gα-binding and -activating (GBA) motif. We combine NMR spectroscopy, computational modelling and biochemistry to map changes in Gα caused by binding of GBA proteins with residue-level resolution. We find that the GBA motif binds to the SwitchII/α3 cleft of Gα and induces changes in the G-1/P-loop and G-2 boxes (involved in phosphate binding), but not in the G-4/G-5 boxes (guanine binding). Our findings reveal that G-protein-binding and activation mechanisms are fundamentally different between GBA proteins and GPCRs, and that GEF-mediated perturbation of nucleotide phosphate binding is sufficient for Gα activation.

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Citations

Nov 15, 2017·Proceedings of the National Academy of Sciences of the United States of America·Anthony LeymeMikel Garcia-Marcos
Jul 4, 2019·Molecular Biology of the Cell·Arthur Marivin, Mikel Garcia-Marcos
Jan 23, 2020·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Dhiraj Srivastava, Nikolai O Artemyev
Jan 18, 2020·The Journal of Biological Chemistry·Arthur MarivinMikel Garcia-Marcos
Jul 30, 2020·Hormone and Metabolic Research = Hormon- Und Stoffwechselforschung = Hormones Et Métabolisme·Anna-Maria WilzCarla Schulz
Apr 6, 2019·The Journal of Cell Biology·Arthur MarivinMikel Garcia-Marcos
Sep 9, 2018·The Journal of Biological Chemistry·Marcin MaziarzMikel Garcia-Marcos
Aug 1, 2019·Proceedings of the National Academy of Sciences of the United States of America·Nicholas A KalogriopoulosIrina Kufareva
Nov 4, 2020·Proceedings of the National Academy of Sciences of the United States of America·Nicholas A KalogriopoulosPradipta Ghosh
Jan 9, 2021·Journal of Structural Biology·Donghee HamKa Young Chung
Mar 7, 2021·The Journal of Biological Chemistry·Jason EarPradipta Ghosh
May 15, 2021·Trends in Pharmacological Sciences·Pradipta Ghosh, Madhubanti Mullick

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Datasets Mentioned

BETA
BAE44387

Methods Mentioned

BETA
nuclear magnetic resonance
co-immunoprecipitation
NMR
nucleotide exchange
peptide array
pull-down
GTPase
our
Infrared Imaging
transfection

Software Mentioned

Molsoft
ICM
Photoshop
Adobe
FoldX
Sparky
Illustrator
PyMOL Molecular Graphics System
Topspin
GIV

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