High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members
Abstract
Regulator of G protein signaling (RGS) proteins are negative regulators of G protein–coupled receptor (GPCR) signaling through their ability to act as GTPase-activating proteins (GAPs) for activated Gα subunits. Members of the RZ subfamily of RGS proteins bind to activated Gαo, Gαz, and Gαi1–3 proteins in the nervous system and thereby inhibit downstream pathways, including those involved in Ca2+-dependent signaling. In contrast to other RGS proteins, little is known about RZ subfamily structure and regulation. In this study, we present the 1.5-Å crystal structure of RGS17, the most complete and highest-resolution structure of an RZ subfamily member to date. RGS17 cocrystallized with Ca2+ bound to conserved positions on the predicted Gα-binding surface of the protein. Using NMR chemical shift perturbations, we confirmed that Ca2+ binds in solution to the same site. Furthermore, RGS17 had greater than 55-fold higher affinity for Ca2+ than for Mg2+. Finally, we found that Ca2+ promotes interactions between RGS17 and activated Gα and decreases the Km for GTP hydrolysis, potentially by altering the binding mechanism between these proteins. Taken together, these findings suggest that Ca2+ positively regulates RGS17, which may represent a general mechanism by which increased Ca2+ concentration promotes the GAP activity of the RZmore »
- Authors:
-
[1]
- Purdue Univ., West Lafayette, IN (United States)
- Univ. of Iowa, Iowa City, IA (United States)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE; American Heart Association; National Institutes of Health (NIH); American Cancer Society
- OSTI Identifier:
- 1569876
- Grant/Contract Number:
- 16SDG29920017; 1R01HL141076-01; 2T32GM008365-26A; IRG-14-190-56
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Volume: 294; Journal Issue: 20; 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; calcium; regulator of G protein signaling (RGS); crystal structure; heterotrimeric G protein; nuclear magnetic resonance (NMR); isothermal titration calorimetry (ITC); cell signaling; GTPase activating protein (GAP); protein crystallization; G protein-coupled receptor (GPCR); RZ subfamily; RGS17
Citation Formats
Sieng, Monita, Hayes, Michael P., O'Brien, Joseph B., Andrew Fowler, C., Houtman, Jon C., Roman, David L., and Lyon, Angeline M. High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members. United States: N. p., 2019.
Web. doi:10.1074/jbc.ra118.006059.
Sieng, Monita, Hayes, Michael P., O'Brien, Joseph B., Andrew Fowler, C., Houtman, Jon C., Roman, David L., & Lyon, Angeline M. High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members. United States. https://doi.org/10.1074/jbc.ra118.006059
Sieng, Monita, Hayes, Michael P., O'Brien, Joseph B., Andrew Fowler, C., Houtman, Jon C., Roman, David L., and Lyon, Angeline M. Tue .
"High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members". United States. https://doi.org/10.1074/jbc.ra118.006059. https://www.osti.gov/servlets/purl/1569876.
@article{osti_1569876,
title = {High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members},
author = {Sieng, Monita and Hayes, Michael P. and O'Brien, Joseph B. and Andrew Fowler, C. and Houtman, Jon C. and Roman, David L. and Lyon, Angeline M.},
abstractNote = {Regulator of G protein signaling (RGS) proteins are negative regulators of G protein–coupled receptor (GPCR) signaling through their ability to act as GTPase-activating proteins (GAPs) for activated Gα subunits. Members of the RZ subfamily of RGS proteins bind to activated Gαo, Gαz, and Gαi1–3 proteins in the nervous system and thereby inhibit downstream pathways, including those involved in Ca2+-dependent signaling. In contrast to other RGS proteins, little is known about RZ subfamily structure and regulation. In this study, we present the 1.5-Å crystal structure of RGS17, the most complete and highest-resolution structure of an RZ subfamily member to date. RGS17 cocrystallized with Ca2+ bound to conserved positions on the predicted Gα-binding surface of the protein. Using NMR chemical shift perturbations, we confirmed that Ca2+ binds in solution to the same site. Furthermore, RGS17 had greater than 55-fold higher affinity for Ca2+ than for Mg2+. Finally, we found that Ca2+ promotes interactions between RGS17 and activated Gα and decreases the Km for GTP hydrolysis, potentially by altering the binding mechanism between these proteins. Taken together, these findings suggest that Ca2+ positively regulates RGS17, which may represent a general mechanism by which increased Ca2+ concentration promotes the GAP activity of the RZ subfamily, leading to RZ-mediated inhibition of Ca2+ signaling.},
doi = {10.1074/jbc.ra118.006059},
journal = {Journal of Biological Chemistry},
number = 20,
volume = 294,
place = {United States},
year = {Tue Apr 02 00:00:00 EDT 2019},
month = {Tue Apr 02 00:00:00 EDT 2019}
}
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