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Title: Structure of the G protein chaperone and guanine nucleotide exchange factor Ric-8A bound to Gαi1

Abstract

Abstract Ric-8A is a cytosolic Guanine Nucleotide exchange Factor (GEF) that activates heterotrimeric G protein alpha subunits (Gα) and serves as an essential Gα chaperone. Mechanisms by which Ric-8A catalyzes these activities, which are stimulated by Casein Kinase II phosphorylation, are unknown. We report the structure of the nanobody-stabilized complex of nucleotide-free Gα bound to phosphorylated Ric-8A at near atomic resolution by cryo-electron microscopy and X-ray crystallography. The mechanism of Ric-8A GEF activity differs considerably from that employed by G protein-coupled receptors at the plasma membrane. Ric-8A engages a specific conformation of Gα at multiple interfaces to form a complex that is stabilized by phosphorylation within a Ric-8A segment that connects two Gα binding sites. The C-terminus of Gα is ejected from its beta sheet core, thereby dismantling the GDP binding site. Ric-8A binds to the exposed Gα beta sheet and switch II to stabilize the nucleotide-free state of Gα.

Authors:
; ORCiD logo; ; ; ; ORCiD logo; ; ORCiD logo; ; ; ; ORCiD logo; ; ORCiD logo
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Advanced Research Projects Agency - Energy (ARPA-E)
Contributing Org.:
Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
OSTI Identifier:
1619535
Alternate Identifier(s):
OSTI ID: 1664417
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Published Article
Journal Name:
Nature Communications
Additional Journal Information:
Journal Name: Nature Communications Journal Volume: 11 Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

McClelland, Levi J., Zhang, Kaiming, Mou, Tung-Chung, Johnston, Jake, Yates-Hansen, Cindee, Li, Shanshan, Thomas, Celestine J., Doukov, Tzanko I., Triest, Sarah, Wohlkonig, Alexandre, Tall, Gregory G., Steyaert, Jan, Chiu, Wah, and Sprang, Stephen R. Structure of the G protein chaperone and guanine nucleotide exchange factor Ric-8A bound to Gαi1. United Kingdom: N. p., 2020. Web. https://doi.org/10.1038/s41467-020-14943-4.
McClelland, Levi J., Zhang, Kaiming, Mou, Tung-Chung, Johnston, Jake, Yates-Hansen, Cindee, Li, Shanshan, Thomas, Celestine J., Doukov, Tzanko I., Triest, Sarah, Wohlkonig, Alexandre, Tall, Gregory G., Steyaert, Jan, Chiu, Wah, & Sprang, Stephen R. Structure of the G protein chaperone and guanine nucleotide exchange factor Ric-8A bound to Gαi1. United Kingdom. https://doi.org/10.1038/s41467-020-14943-4
McClelland, Levi J., Zhang, Kaiming, Mou, Tung-Chung, Johnston, Jake, Yates-Hansen, Cindee, Li, Shanshan, Thomas, Celestine J., Doukov, Tzanko I., Triest, Sarah, Wohlkonig, Alexandre, Tall, Gregory G., Steyaert, Jan, Chiu, Wah, and Sprang, Stephen R. Wed . "Structure of the G protein chaperone and guanine nucleotide exchange factor Ric-8A bound to Gαi1". United Kingdom. https://doi.org/10.1038/s41467-020-14943-4.
@article{osti_1619535,
title = {Structure of the G protein chaperone and guanine nucleotide exchange factor Ric-8A bound to Gαi1},
author = {McClelland, Levi J. and Zhang, Kaiming and Mou, Tung-Chung and Johnston, Jake and Yates-Hansen, Cindee and Li, Shanshan and Thomas, Celestine J. and Doukov, Tzanko I. and Triest, Sarah and Wohlkonig, Alexandre and Tall, Gregory G. and Steyaert, Jan and Chiu, Wah and Sprang, Stephen R.},
abstractNote = {Abstract Ric-8A is a cytosolic Guanine Nucleotide exchange Factor (GEF) that activates heterotrimeric G protein alpha subunits (Gα) and serves as an essential Gα chaperone. Mechanisms by which Ric-8A catalyzes these activities, which are stimulated by Casein Kinase II phosphorylation, are unknown. We report the structure of the nanobody-stabilized complex of nucleotide-free Gα bound to phosphorylated Ric-8A at near atomic resolution by cryo-electron microscopy and X-ray crystallography. The mechanism of Ric-8A GEF activity differs considerably from that employed by G protein-coupled receptors at the plasma membrane. Ric-8A engages a specific conformation of Gα at multiple interfaces to form a complex that is stabilized by phosphorylation within a Ric-8A segment that connects two Gα binding sites. The C-terminus of Gα is ejected from its beta sheet core, thereby dismantling the GDP binding site. Ric-8A binds to the exposed Gα beta sheet and switch II to stabilize the nucleotide-free state of Gα.},
doi = {10.1038/s41467-020-14943-4},
journal = {Nature Communications},
number = 1,
volume = 11,
place = {United Kingdom},
year = {2020},
month = {2}
}

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