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Title: Structure of the Regulator of G Protein Signaling 8 (RGS8)-Gα q Complex: Molecular Basis for Gα Selectivity

Abstract

Regulator of G protein signaling (RGS) proteins interact with activated Gα subunits via their RGS domains and accelerate the hydrolysis of GTP. Although the R4 subfamily of RGS proteins generally accepts both Gα i/o and Gα q/11 subunits as substrates, the R7 and R12 subfamilies select against Gα q/11. In contrast, only one RGS protein, RGS2, is known to be selective for Gαq/11. The molecular basis for this selectivity is not clear. Previously, the crystal structure of RGS2 in complex with Gα q revealed a non-canonical interaction that could be due to interfacial differences imposed by RGS2, the Gα subunit, or both. To resolve this ambiguity, the 2.6 Å crystal structure of RGS8, an R4 subfamily member, was determined in complex with Gα q. RGS8 adopts the same pose on Gα q as it does when bound to Gα i3, indicating that the non-canonical interaction of RGS2 with Gα q is due to unique features of RGS2. Here, based on the RGS8-Gα q structure, residues in RGS8 that contact a unique α-helical domain loop of Gα q were converted to those typically found in R12 subfamily members, and the reverse substitutions were introduced into RGS10, an R12 subfamily member. Althoughmore » these substitutions perturbed their ability to stimulate GTP hydrolysis, they did not reverse selectivity. Instead, selectivity for Gα q seems more likely determined by whether strong contacts can be maintained between α6 of the RGS domain and Switch III of Gα q, regions of high sequence and conformational diversity in both protein families.« less

Authors:
 [1];  [1];  [1]
  1. Univ. of Michigan, Ann Arbor, MI (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Scientific User Facilities Division; National Inst. of Health; Michigan Economic Development Corporation; Michigan Technology Tri-Corridor
OSTI Identifier:
1243106
Grant/Contract Number:  
HL086865; HL122416; T32GM008270; AC02-06CH11357; 085P1000817
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 291; Journal Issue: 10; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; crystal structure; GTPase-activating protein (GAP); heterotrimeric G protein; regulator of G protein signaling (RGS); structure-function; X-ray crystallography; Gq; RGS10; RGS8; selectivity; cell signaling; protein structure; cardiovascular disease; protein complex; GTPase activation assays; Gαq

Citation Formats

Taylor, Veronica G., Bommarito, Paige A., and Tesmer, John J. G. Structure of the Regulator of G Protein Signaling 8 (RGS8)-Gαq Complex: Molecular Basis for Gα Selectivity. United States: N. p., 2016. Web. doi:10.1074/jbc.M115.712075.
Taylor, Veronica G., Bommarito, Paige A., & Tesmer, John J. G. Structure of the Regulator of G Protein Signaling 8 (RGS8)-Gαq Complex: Molecular Basis for Gα Selectivity. United States. doi:10.1074/jbc.M115.712075.
Taylor, Veronica G., Bommarito, Paige A., and Tesmer, John J. G. Mon . "Structure of the Regulator of G Protein Signaling 8 (RGS8)-Gαq Complex: Molecular Basis for Gα Selectivity". United States. doi:10.1074/jbc.M115.712075. https://www.osti.gov/servlets/purl/1243106.
@article{osti_1243106,
title = {Structure of the Regulator of G Protein Signaling 8 (RGS8)-Gαq Complex: Molecular Basis for Gα Selectivity},
author = {Taylor, Veronica G. and Bommarito, Paige A. and Tesmer, John J. G.},
abstractNote = {Regulator of G protein signaling (RGS) proteins interact with activated Gα subunits via their RGS domains and accelerate the hydrolysis of GTP. Although the R4 subfamily of RGS proteins generally accepts both Gαi/o and Gαq/11 subunits as substrates, the R7 and R12 subfamilies select against Gαq/11. In contrast, only one RGS protein, RGS2, is known to be selective for Gαq/11. The molecular basis for this selectivity is not clear. Previously, the crystal structure of RGS2 in complex with Gαq revealed a non-canonical interaction that could be due to interfacial differences imposed by RGS2, the Gα subunit, or both. To resolve this ambiguity, the 2.6 Å crystal structure of RGS8, an R4 subfamily member, was determined in complex with Gαq. RGS8 adopts the same pose on Gαq as it does when bound to Gαi3, indicating that the non-canonical interaction of RGS2 with Gαq is due to unique features of RGS2. Here, based on the RGS8-Gαq structure, residues in RGS8 that contact a unique α-helical domain loop of Gαq were converted to those typically found in R12 subfamily members, and the reverse substitutions were introduced into RGS10, an R12 subfamily member. Although these substitutions perturbed their ability to stimulate GTP hydrolysis, they did not reverse selectivity. Instead, selectivity for Gαq seems more likely determined by whether strong contacts can be maintained between α6 of the RGS domain and Switch III of Gαq, regions of high sequence and conformational diversity in both protein families.},
doi = {10.1074/jbc.M115.712075},
journal = {Journal of Biological Chemistry},
issn = {0021-9258},
number = 10,
volume = 291,
place = {United States},
year = {2016},
month = {1}
}

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Works referenced in this record:

Transducin GTPase provides for rapid quenching of the cGMP cascade in rod outer segments
journal, July 1989


A Single Mutation Asp 229 → Ser Confers upon G s α the Ability To Interact with Regulators of G Protein Signaling
journal, September 1998

  • Natochin, Michael; Artemyev, Nikolai O.
  • Biochemistry, Vol. 37, Issue 39
  • DOI: 10.1021/bi981155a

Structure of RGS4 Bound to AlF4−-Activated Giα1: Stabilization of the Transition State for GTP Hydrolysis
journal, April 1997


Overview of the CCP 4 suite and current developments
journal, March 2011

  • Winn, Martyn D.; Ballard, Charles C.; Cowtan, Kevin D.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4
  • DOI: 10.1107/S0907444910045749

Selective Inhibition of α 1A -Adrenergic Receptor Signaling by RGS2 Association with the Receptor Third Intracellular Loop
journal, May 2005

  • Hague, Chris; Bernstein, Leah S.; Ramineni, Suneela
  • Journal of Biological Chemistry, Vol. 280, Issue 29
  • DOI: 10.1074/jbc.M502365200

Identification of RGS2 and Type V Adenylyl Cyclase Interaction Sites
journal, February 2003

  • Salim, Samina; Sinnarajah, Srikumar; Kehrl, John H.
  • Journal of Biological Chemistry, Vol. 278, Issue 18
  • DOI: 10.1074/jbc.M210663200

Modulation of the Affinity and Selectivity of RGS Protein Interaction with G α Subunits by a Conserved Asparagine/Serine Residue
journal, June 1999

  • Posner, Bruce A.; Mukhopadhyay, Suchetana; Tesmer, John J.
  • Biochemistry, Vol. 38, Issue 24
  • DOI: 10.1021/bi9906367

Mechanism of GTP hydrolysis by G-protein alpha subunits.
journal, October 1994

  • Kleuss, C.; Raw, A. S.; Lee, E.
  • Proceedings of the National Academy of Sciences, Vol. 91, Issue 21
  • DOI: 10.1073/pnas.91.21.9828

Integrating energy calculations with functional assays to decipher the specificity of G protein–RGS protein interactions
journal, June 2011

  • Kosloff, Mickey; Travis, Amanda M.; Bosch, Dustin E.
  • Nature Structural & Molecular Biology, Vol. 18, Issue 7
  • DOI: 10.1038/nsmb.2068

Structural Determinants of G-protein α Subunit Selectivity by Regulator of G-protein Signaling 2 (RGS2)
journal, May 2009

  • Kimple, Adam J.; Soundararajan, Meera; Hutsell, Stephanie Q.
  • Journal of Biological Chemistry, Vol. 284, Issue 29
  • DOI: 10.1074/jbc.M109.024711

GTPase-Activating Proteins for Heterotrimeric G Proteins: Regulators of G Protein Signaling (RGS) and RGS-Like Proteins
journal, June 2000


Structural diversity in the RGS domain and its interaction with heterotrimeric G protein  -subunits
journal, April 2008

  • Soundararajan, M.; Willard, F. S.; Kimple, A. J.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 17
  • DOI: 10.1073/pnas.0801508105

Characterization of the GRK2 Binding Site of Gα q
journal, October 2004

  • Day, Peter W.; Tesmer, John J. G.; Sterne-Marr, Rachel
  • Journal of Biological Chemistry, Vol. 279, Issue 51
  • DOI: 10.1074/jbc.M401438200

The Regulator of G Protein Signaling Family
journal, April 2000


Assembly of High Order Gα q -Effector Complexes with RGS Proteins
journal, October 2008

  • Shankaranarayanan, Aruna; Thal, David M.; Tesmer, Valerie M.
  • Journal of Biological Chemistry, Vol. 283, Issue 50
  • DOI: 10.1074/jbc.M805860200

RGS family members: GTPase-activating proteins for heterotrimeric G-protein α-subunits
journal, September 1996

  • Watson, Ned; Linder, Maurine E.; Druey, Kirk M.
  • Nature, Vol. 383, Issue 6596
  • DOI: 10.1038/383172a0

Hypertension and prolonged vasoconstrictor signaling in RGS2-deficient mice
journal, February 2003

  • Heximer, Scott P.; Knutsen, Russell H.; Sun, Xiaoguang
  • Journal of Clinical Investigation, Vol. 111, Issue 4
  • DOI: 10.1172/JCI200315598

Regulator of G-protein signaling-2 mediates vascular smooth muscle relaxation and blood pressure
journal, November 2003

  • Tang, Mary; Wang, Guang; Lu, Ping
  • Nature Medicine, Vol. 9, Issue 12
  • DOI: 10.1038/nm958

Heterotrimeric G protein activation by G-protein-coupled receptors
journal, January 2008

  • Oldham, William M.; Hamm, Heidi E.
  • Nature Reviews Molecular Cell Biology, Vol. 9, Issue 1
  • DOI: 10.1038/nrm2299

Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions
journal, November 2004

  • Krissinel, E.; Henrick, K.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12
  • DOI: 10.1107/S0907444904026460

The N-terminal Domain of RGS4 Confers Receptor-selective Inhibition of G Protein Signaling
journal, December 1998

  • Zeng, Weizhang; Xu, Xin; Popov, Serguei
  • Journal of Biological Chemistry, Vol. 273, Issue 52
  • DOI: 10.1074/jbc.273.52.34687

Inhibition of G-protein signaling by dominant gain-of-function mutations in Sst2p, a pheromone desensitization factor in Saccharomyces cerevisiae.
journal, July 1995

  • Dohlman, H. G.; Apaniesk, D.; Chen, Y.
  • Molecular and Cellular Biology, Vol. 15, Issue 7
  • DOI: 10.1128/MCB.15.7.3635

Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega
journal, January 2011

  • Sievers, Fabian; Wilm, Andreas; Dineen, David
  • Molecular Systems Biology, Vol. 7, Issue 1
  • DOI: 10.1038/msb.2011.75

Increased Expression of Regulator of G Protein Signaling-2 (RGS-2) in Bartter’s/Gitelman’s Syndrome. A Role in the Control of Vascular Tone and Implication for Hypertension
journal, August 2004

  • Calò, Lorenzo A.; Pagnin, Elisa; Davis, Paul A.
  • The Journal of Clinical Endocrinology & Metabolism, Vol. 89, Issue 8
  • DOI: 10.1210/jc.2004-0498

The α-Helical Domain of Gα t Determines Specific Interaction with Regulator of G Protein Signaling 9
journal, March 1999

  • Skiba, Nikolai P.; Yang, Chii-Shen; Huang, Tao
  • Journal of Biological Chemistry, Vol. 274, Issue 13
  • DOI: 10.1074/jbc.274.13.8770

Structural and Functional Analysis of the Regulator of G Protein Signaling 2-Gαq Complex
journal, March 2013


RGS17/RGSZ2, a Novel Regulator of G i/o , G z , and G q Signaling
journal, April 2004

  • Mao, Helen; Zhao, Qingshi; Daigle, Mireille
  • Journal of Biological Chemistry, Vol. 279, Issue 25
  • DOI: 10.1074/jbc.M401800200

Mu opioid receptor activation enhances regulator of G protein signaling 4 association with the mu opioid receptor/G protein complex in a GTP-dependent manner
journal, July 2015

  • Santhappan, Rema; Crowder, Alicia Tamara; Gouty, Shawn
  • Journal of Neurochemistry, Vol. 135, Issue 1
  • DOI: 10.1111/jnc.13222

The RGS2 Gene Product from a Candidate Hypertension Allele Shows Decreased Plasma Membrane Association and Inhibition of G q
journal, January 2008

  • Gu, Steven; Tirgari, Sam; Heximer, Scott P.
  • Molecular Pharmacology, Vol. 73, Issue 4
  • DOI: 10.1124/mol.107.044214

G Protein Selectivity Is a Determinant of RGS2 Function
journal, November 1999

  • Heximer, Scott P.; Srinivasa, Sreesha P.; Bernstein, Leah S.
  • Journal of Biological Chemistry, Vol. 274, Issue 48
  • DOI: 10.1074/jbc.274.48.34253

Inference of Macromolecular Assemblies from Crystalline State
journal, September 2007