Mapping the Putative G Protein-coupled Receptor (GPCR) Docking Site on GPCR Kinase 2

Beautrait, Alexandre; Michalski, Kevin; Lopez, T.; Mannix, Katelynn M.; McDonald, Devin; Cutter, Amber R.; Medina, Christopher B.; Hebert, Aaron; Francis, Charnelle; Bouvier, Michel; Tesmer, John J.G.; Sterne‐Marr, Rachel

doi:10.1074/jbc.m114.593178

Cited by 35 publications

(42 citation statements)

References 54 publications

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“…To understand how GRK2 may function in NIH/3T3 cells, we took advantage of previous studies that have mapped critical amino acid residues in GRK2 that are required for kinase activity, membrane recruitment, interaction with GPCRs, and binding to G protein subunits (32–35) (Fig. 3A).…”

Section: Resultsmentioning

confidence: 99%

G protein–coupled receptors control the sensitivity of cells to the morphogen Sonic Hedgehog

et al. 2018

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The morphogen Sonic Hedgehog (SHH) patterns tissues during development by directing cell fates in a concentration-dependent manner. The SHH signal is transmitted across the membrane of target cells by the heptahelical transmembrane protein Smoothened (SMO), which activates the GLI family of transcription factors through a mechanism that is undefined in vertebrates. Using CRISPR-edited null alleles and small molecule inhibitors, we systematically analyzed the epistatic interactions between SMO and three proteins implicated in SMO signaling: the heterotrimeric G-protein subunit GαS, G protein–coupled receptor kinase 2 (GRK2), and the GαS-coupled receptor GPR161. Our experiments uncovered a signaling mechanism that modifies the sensitivity of target cells to SHH and consequently changes the shape of the SHH dose-response curve. In both fibroblasts and spinal neural progenitors, the loss of GPR161, previously implicated as an inhibitor of basal SHH signaling, increased the sensitivity of target cells across the entire spectrum of SHH concentrations. Surprisingly, GRK2, thought to function by antagonizing GPR161, and Gαs, which is activated by GPR161, influenced SHH signaling even in cells lacking GPR161. We propose that the sensitivity of target cells to Hedgehog (Hh) morphogens, and the consequent effects on gene expression and differentiation outcomes, can be controlled by signals from G-protein coupled receptors that converge on Gαs and Protein Kinase A.

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Section: Resultsmentioning

confidence: 99%

G protein–coupled receptors control the sensitivity of cells to the morphogen Sonic Hedgehog

et al. 2018

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“…Often, the key contribution of GRKs in mediating the switch between G protein and barr engagement is overlooked when screening for or assessing biased drugs. Structural work investigating the mechanism of GRK5 activation by GPCRs and investigations of GRK2 activation (23,24) indicate that GRK activation is a key feature of GPCR signal transduction. Therefore, elucidating this and other mechanisms of selective G protein or barr pathway engagement will inform future efforts to design functionally selective GPCR therapeutics.…”

Section: Discussionmentioning

confidence: 99%

“…A notable study by Beautrait et al (23), showed both G protein-and GPCR-mediated components of GRK2 recruitment and mapped the residues on the N-terminus of GRK2 involved in mediating recruitment and/or phosphorylation. Indeed, recent structural work at the b2AR and GRK5 demonstrates that GRK5 activation occurs via a rearrangement of GRK5's RH/catalytic domain, which facilitates interaction between the GRK's catalytic domain and the GPCR's intracellular loops (24).…”

Section: Introductionmentioning

confidence: 99%

The dopamine D2 receptor can directly recruit and activate GRK2 without G protein activation

Pack

Orlen

Ray

et al. 2018

Journal of Biological Chemistry

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The dopamine D2 receptor (D2R) is a G protein-coupled receptor (GPCR) that is critical for many central nervous system functions. The D2R carries out these functions by signaling through two transducers: G proteins and b-arrestins (barrs). Selectively engaging either the G protein or barr pathway may be a way to improve drugs targeting GPCRs. The current model of GPCR signal transduction posits a chain of events where G protein activation ultimately leads to barr recruitment. GPCR kinases (GRKs), which are regulated by G proteins and whose kinase action facilitates barr recruitment, bridge these pathways. Therefore barr recruitment appears to be intimately tied to G protein activation via GRKs. Here, we sought to understand how GRK2 action at the D2R would be disrupted when G protein activation is eliminated and the effect of this on barr recruitment. We used two recently developed biased D2R mutants that can preferentially interact either with G proteins or barrs as well as a barr-biased D2R ligand, UNC9994. With these functionally selective tools, we investigated the mechanism whereby the barr-preferring D2R achieves barr pathway activation in the complete absence of G protein activation. We describe how direct, G protein-independent recruitment of GRK2 drives interactions at the barr-preferring D2R and also contributes to barr recruitment at the WT D2R. Additionally, we found an additive interaction between the barr-preferring D2R mutant and UNC9994. These results reveal that the D2R can directly recruit GRK2 without G protein activation and that this mechanism may have relevance to achieving barr-biased signaling.

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“…lower affinity binding to the kinase active site (46). The C-tail is thought to play a role in kinase domain closure and activation (47)(48)(49). Full ordering of the C-tail is seen in chain A of the GRK4␣ structure (Fig.…”

Section: Discussionmentioning

confidence: 99%

Structure and Function of the Hypertension Variant A486V of G Protein-coupled Receptor Kinase 4

Allen

Parthasarathy

Darke

et al. 2015

Journal of Biological Chemistry

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Background: GRK4 mediates GPCR phosphorylation and is implicated in hypertension. Results: The crystal structure of human GRK4␣ A486V is presented. Phosphorylation assays highlight kinetic differences between wild-type GRK4␣ and GRK4␣ A486V. Conclusion: GRK4␣ has unusual features that help explain the effects of GRK4 mutations and GRK4 biology. Significance: This work provides a structural basis for GRK4 function.

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Mapping the Putative G Protein-coupled Receptor (GPCR) Docking Site on GPCR Kinase 2

Cited by 35 publications

References 54 publications

G protein–coupled receptors control the sensitivity of cells to the morphogen Sonic Hedgehog

G protein–coupled receptors control the sensitivity of cells to the morphogen Sonic Hedgehog

The dopamine D2 receptor can directly recruit and activate GRK2 without G protein activation

Structure and Function of the Hypertension Variant A486V of G Protein-coupled Receptor Kinase 4

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