Structural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor

Dhiraj Srivastava; Lokesh Gakhar; Nikolai O Artemyev

doi:10.1038/s41467-019-11088-x

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Structural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor

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Structural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor

Dhiraj Srivastava, Lokesh Gakhar and Nikolai O Artemyev

Nature communications, Vol.10(1), pp.3084-14

07/12/2019

DOI: 10.1038/s41467-019-11088-x

PMCID: PMC6625990

PMID: 31300652

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Abstract

Resistance to inhibitors of cholinesterase 8A (Ric8A) is an essential regulator of G protein α-subunits (Gα), acting as a guanine nucleotide exchange factor and a chaperone. We report two crystal structures of Ric8A, one in the apo form and the other in complex with a tagged C-terminal fragment of Gα. These structures reveal two principal domains of Ric8A: an armadillo-fold core and a flexible C-terminal tail. Additionally, they show that the Gα C-terminus binds to a highly-conserved patch on the concave surface of the Ric8A armadillo-domain, with selectivity determinants residing in the Gα sequence. Biochemical analysis shows that the Ric8A C-terminal tail is critical for its stability and function. A model of the Ric8A/Gα complex derived from crosslinking mass spectrometry and molecular dynamics simulations suggests that the Ric8A C-terminal tail helps organize the GTP-binding site of Gα. This study lays the groundwork for understanding Ric8A function at the molecular level.

Recombinant Proteins - metabolism

Guanine Nucleotide Exchange Factors - genetics

GTP-Binding Protein alpha Subunits - metabolism

Molecular Chaperones - metabolism

Mutagenesis, Site-Directed

Recombinant Proteins - ultrastructure

Molecular Chaperones - genetics

Scattering, Small Angle

Guanine Nucleotide Exchange Factors - ultrastructure

Recombinant Proteins - genetics

Armadillo Domain Proteins - genetics

Molecular Dynamics Simulation

Animals

Cattle

Guanine Nucleotide Exchange Factors - metabolism

X-Ray Diffraction

Armadillo Domain Proteins - metabolism

Armadillo Domain Proteins - ultrastructure

Molecular Chaperones - ultrastructure

Details

Title: Subtitle: Structural underpinnings of Ric8A function as a G-protein α-subunit chaperone and guanine-nucleotide exchange factor
Creators: Dhiraj Srivastava - Department of Molecular Physiology and Biophysics, University of Iowa Carver College of Medicine, Iowa City, IA, 52242, USA
Lokesh Gakhar - Protein Crystallography Facility, University of Iowa Carver College of Medicine, Iowa City, IA, 52242, USA
Nikolai O Artemyev - Department of Ophthalmology and Visual Sciences, University of Iowa Carver College of Medicine, Iowa City, IA, 52242, USA. nikolai-artemyev@uiowa.edu
Resource Type: Journal article
Publication Details: Nature communications, Vol.10(1), pp.3084-14
DOI: 10.1038/s41467-019-11088-x
PMID: 31300652
PMCID: PMC6625990
NLM abbreviation: Nat Commun
ISSN: 2041-1723
eISSN: 2041-1723
Publisher: England
Grant note: P41 GM103622 / NIGMS NIH HHS RO1 EY-12682 / U.S. Department of Health & Human Services | NIH | National Eye Institute (NEI) R01 EY012682 / NEI NIH HHS S10 OD018090 / NIH HHS
Language: English
Date published: 07/12/2019
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Biochemistry and Molecular Biology; Medicine Administration; Ophthalmology and Visual Sciences
Record Identifier: 9984070649502771

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