High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members

Sieng, Monita; Hayes, Michael P.; O'Brien, Joseph B.; Andrew Fowler, C.; Houtman, Jon C.; Roman, David L.; Lyon, Angeline M.

doi:10.1074/jbc.ra118.006059

Title: High-resolution structure of RGS17 suggests a role for Ca²⁺ in promoting the GTPase-activating protein activity by RZ subfamily members

Journal Article · Tue Apr 02 00:00:00 EDT 2019 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.ra118.006059· OSTI ID:1569876

Sieng, Monita ^[1]; Hayes, Michael P. ^[2]; O'Brien, Joseph B. ^[2]; Andrew Fowler, C. ^[2]; Houtman, Jon C. ^[2]; Roman, David L. ^[2];

^[1]

Purdue Univ., West Lafayette, IN (United States)
Univ. of Iowa, Iowa City, IA (United States)

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 Gα_o, Gα_z, and Gα_i1–3 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 K_m 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.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE; American Heart Association; National Institutes of Health (NIH); American Cancer Society

Grant/Contract Number:: 16SDG29920017; 1R01HL141076-01; 2T32GM008365-26A; IRG-14-190-56

OSTI ID:: 1569876

Journal Information:: Journal of Biological Chemistry, Vol. 294, Issue 20; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 2 works

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Related Subjects

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

Title: High-resolution structure of RGS17 suggests a role for Ca2+ in promoting the GTPase-activating protein activity by RZ subfamily members

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Title: High-resolution structure of RGS17 suggests a role for Ca²⁺ in promoting the GTPase-activating protein activity by RZ subfamily members