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Phosphorylation-independent desensitization of metabotropic glutamate receptor 5 by G protein-coupled receptor kinase 2 in HEK 293 cells

  • Molecular Biology of the Cell
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Abstract

The metabotropic glutamate receptor 5 (mGluR5) is one of the important excitatory neurotransmitter receptors in the central nervous system, and its desensitization by G protein-coupled receptor kinases (GRKs) plays an important role in neuron protection against receptor overstimulation. It is reported that GRK2 could down-regulate the mGluR5 signaling in both HEK 293 cells and neurons. However, whether GRK2-mediated mGluR5 desensitization is phosphorylation dependent remains controversial. Here, we demonstrated that the signal intensity and kinetics of mGluR5 desensitization was inhibited or changed by GRK2 in HEK 293 cells. By using the catalytically inactive GRK2 mutant K220R, and the receptor mutants that lack potential phosphorylation sites in the C-terminal tail, we demonstrated that the GRK2-mediated mGluR5 desensitization was phosphorylation-independent. Furthermore, overexpression of an N-terminal regulator of G protein signaling (RGS) homology (RH) domain of GRK2 was sufficient to attenuate the mGluR5 signaling, whereas the expression of GRK2 D110A mutant devoid in Gαq binding was unable to inhibit mGluR5 signaling. In summary, this study provides evidence that GRK2 mediates phosphorylationindependent mGluR5 desensitization via the interaction between the RGS domain and Gαq in HEK 293 cells.

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Abbreviations

Gαq:

α subunit of G protein

GPCRs:

G protein coupled receptors

GRK:

G protein-coupled receptor kinase

InsP:

inositol-1,4,5-triphosphate

ELISA:

enzyme-linked immunosorbent assay

mGluR5:

metabotropic glutamate receptor 5

GIRK:

G-protein coupled inward rectifier potassium channel

PH domain:

pleckstrin homology domain

RGS:

regulator of G protein signaling

RH domain:

RGS homology domain

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Authors and Affiliations

  1. Sino-France Laboratory for Drug Screening, Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, Hubei, China

    Z. Zhang, L. Xue, H. Guo, Y. Li, H. Ding & S. Huang

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  1. Z. Zhang
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  2. L. Xue
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  3. H. Guo
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  4. Y. Li
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  5. H. Ding
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  6. S. Huang
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Correspondence to S. Huang.

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Zhang, Z., Xue, L., Guo, H. et al. Phosphorylation-independent desensitization of metabotropic glutamate receptor 5 by G protein-coupled receptor kinase 2 in HEK 293 cells. Mol Biol 47, 123–130 (2013). https://doi.org/10.1134/S0026893313010160

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  • DOI: https://doi.org/10.1134/S0026893313010160

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