Logo image
Large-scale conformational rearrangement of the α5-helix of Gα subunits in complex with the guanine nucleotide exchange factor Ric8A
Journal article   Open access   Peer reviewed

Large-scale conformational rearrangement of the α5-helix of Gα subunits in complex with the guanine nucleotide exchange factor Ric8A

Dhiraj Srivastava and Nikolai O Artemyev
The Journal of biological chemistry, Vol.294(47), pp.17875-17882
11/22/2019
DOI: 10.1074/jbc.AC119.011135
PMCID: PMC6879328
PMID: 31624147
url
https://doi.org/10.1074/jbc.AC119.011135View
Published (Version of record) Open Access

Abstract

Resistance to inhibitors of cholinesterase 8A (Ric8A) protein is an important G protein-coupled receptor (GPCR)-independent regulator of G protein α-subunits (Gα), acting as a guanine nucleotide exchange factor (GEF) and a chaperone. Insights into the complex between Ric8A and Gα hold the key to understanding the mechanisms underlying noncanonical activation of G-protein signaling as well as the folding of nascent Gα proteins. Here, we examined the structure of the complex of Ric8A with minimized Gα (miniGα ) in solution by small-angle X-ray scattering (SAXS) and exploited the scattering profile in modeling of the Ric8A/miniGα complex by steered molecular dynamics (SMD) simulations. A small set of models of the complex featured minimal clash scores, excellent agreement with the experimental SAXS data, and a large-scale rearrangement of the signal-transducing α5-helix of Gα away from its β-sheet core. The resulting interface involved the Gα α5-helix bound to the concave surface of Ric8A and the Gα β-sheet that wraps around the C-terminal part of the Ric8A armadillo domain, leading to a severe disruption of the GDP-binding site. Further modeling of the flexible C-terminal tail of Ric8A indicated that it interacts with the effector surface of Gα. This smaller interface may enable the Ric8A-bound Gα to interact with GTP. The two-interface interaction with Gα described here distinguishes Ric8A from GPCRs and non-GPCR regulators of G-protein signaling.
Animals GTP-Binding Protein alpha Subunits - metabolism Protein Structure, Secondary Cattle GTP-Binding Protein alpha Subunits - chemistry Guanine Nucleotide Exchange Factors - metabolism X-Ray Diffraction Scattering, Small Angle Static Electricity Molecular Dynamics Simulation Guanine Nucleotide Exchange Factors - chemistry

Details

Logo image