Distinct phosphorylation sites on the ghrelin receptor, GHSR1a, establish a code that determines the functions of ß-arrestins

Bouzo-Lorenzo, Monica; Santo-Zas, Icía; Lodeiro, María F.; Nogueiras, Rubén; Casanueva, Felipe F.; Castro, Marián; Pazos, Yolanda; Tobin, Andrew B.; Butcher, Adrian J.; Camiña, Jesús P.

doi:10.1038/srep22495

Cited by 40 publications

(55 citation statements)

References 33 publications

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“…Indeed, the position of phosphorylation sites within the receptor C-terminus or intracellular loop regions appears to be key to regulating arrestin recruitment and receptor function. This certainly appears to be the case for PTH1R as phosphorylation sites within cluster 1 have a smaller impact on arrestin recruitment, suggesting that phosphorylation within cluster 1 may perform a modulatory or stabilizing role possibly fine-tuning arrestin–receptor interactions as has been suggested for other receptors [32]. In contrast, phosphorylation within cluster 2 is responsible for establishing the high-affinity receptor–arrestin interaction.…”

Section: Discussionmentioning

confidence: 93%

“…Our recent studies on the M 3 -muscarinic receptor [31], GHSR1a [32] and GPR120 [26], had indicated that distinct phosphorylation sites, or patterns of receptor phosphorylation, could mediate differential signaling outcomes, including differences in receptor–arrestin interaction. Hence, we examined whether phosphorylation at clusters 1 and 2 of the PTH1R contributed equally to receptor/arrestin interaction.…”

Section: Resultsmentioning

confidence: 99%

“…Patterns of receptor phosphorylation have been determined for receptors such as FFAR4, the ghrelin receptor (GHSR1a), chemokine receptor CXCR4, the β 2 AR and the M 3 -muscarinic receptor [26,31,32,48], and it would appear at least in these instances that receptor phosphorylation represents more than just the addition of bulk negative charge to a certain receptor domain. Indeed, the position of phosphorylation sites within the receptor C-terminus or intracellular loop regions appears to be key to regulating arrestin recruitment and receptor function.…”

Section: Discussionmentioning

confidence: 99%

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Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Zindel

Engel

Bottrill

et al. 2016

Biochemical Journal

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The parathyroid hormone receptor 1 (PTH1R) is a member of family B of G-protein-coupled receptors (GPCRs), predominantly expressed in bone and kidney where it modulates extracellular Ca2+ homeostasis and bone turnover. It is well established that phosphorylation of GPCRs constitutes a key event in regulating receptor function by promoting arrestin recruitment and coupling to G-protein-independent signaling pathways. Mapping phosphorylation sites on PTH1R would provide insights into how phosphorylation at specific sites regulates cell signaling responses and also open the possibility of developing therapeutic agents that could target specific receptor functions. Here, we have used mass spectrometry to identify nine sites of phosphorylation in the C-terminal tail of PTH1R. Mutational analysis revealed identified two clusters of serine and threonine residues (Ser489–Ser495 and Ser501–Thr506) specifically responsible for the majority of PTH(1–34)-induced receptor phosphorylation. Mutation of these residues to alanine did not affect negatively on the ability of the receptor to couple to G-proteins or activate extracellular-signal-regulated kinase 1/2. Using fluorescence resonance energy transfer and bioluminescence resonance energy transfer to monitor PTH(1–34)-induced interaction of PTH1R with arrestin3, we show that the first cluster Ser489–Ser495 and the second cluster Ser501–Thr506 operated in concert to mediate both the efficacy and potency of ligand-induced arrestin3 recruitment. We further demonstrate that Ser503 and Thr504 in the second cluster are responsible for 70% of arrestin3 recruitment and are key determinants for interaction of arrestin with the receptor. Our data are consistent with the hypothesis that the pattern of C-terminal tail phosphorylation on PTH1R may determine the signaling outcome following receptor activation.

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Section: Discussionmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Zindel

Engel

Bottrill

et al. 2016

Biochemical Journal

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“…Considerable effort has been made during recent years to define this for a variety of receptors, mainly using partial affinity purification and MS (see some recent examples: Alfonzo-Méndez, Alcántara-Hernández, & García-Sáinz, 2017;Alvarez-Curto et al, 2016;Bouzo-Lorenzo et al, 2016;Bradley et al, 2016;Butcher et al, 2011;Butcher et al, 2014;Prihandoko et al, 2016;Zindel et al, 2016). Alfonzo .…”

Section: Regulation Of α 1 -Adrenoceptors By Phosphorylationmentioning

confidence: 99%

Updates in the function and regulation of α₁‐adrenoceptors

Akinaga

García‐Sáinz

Pupo

2019

British J Pharmacology

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α1‐Adrenoceptors are seven transmembrane domain GPCRs involved in numerous physiological functions controlled by the endogenous catecholamines, noradrenaline and adrenaline, and targeted by drugs useful in therapeutics. Three separate genes, whose products are named α1A‐, α1B‐, and α1D‐ adrenoceptors, encode these receptors. Although the existence of multiple α1‐adrenoceptors has been acknowledged for almost 25 years, the specific functions regulated by each subtype are still largely unknown. Despite the limited comprehension, the identification of a single class of subtype‐selective ligands for the α1A‐ adrenoceptors, the so‐called α‐blockers for prostate dysfunction, has led to major improvement in therapeutics, demonstrating the need for continued efforts in the field. This review article surveys the tissue distribution of the three α1‐adrenoceptor subtypes in the cardiovascular system, genitourinary system, and CNS, highlighting the functions already identified as mediated by the predominant activation of specific subtypes. In addition, this review covers the recent advances in the understanding of the molecular mechanisms involved in the regulation of each of the α1‐adrenoceptor subtypes by phosphorylation and interaction with proteins involved in their desensitization and internalization. Linked Articles This article is part of a themed section on Adrenoceptors—New Roles for Old Players. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v176.14/issuetoc

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“…Mutagenesis of the S/T residues involved in arrestin binding also prevented co-internalization of the PTH1R with arrestin upon agonist stimulation. Patterns of GPCR phosphorylation were also determined for a variety of GPCR such as gherlin receptor, chemokine receptor CXCR4 and M3 muscarinic receptor [37-42]. …”

Section: Role Of Agonist-promoted Phosphorylation “Barcoding” In Rmentioning

confidence: 99%

Barcoding of GPCR trafficking and signaling through the various trafficking roadmaps by compartmentalized signaling networks

Bahouth

Nooh

2017

Cellular Signalling

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Proper signaling by G protein coupled receptors (GPCR) is dependent on the specific repertoire of transducing, enzymatic and regulatory kinases and phosphatases that shape its signaling output. Activation and signaling of the GPCR through its cognate G protein is impacted by G protein-coupled receptor kinase (GRK)-imprinted “barcodes” that recruit ß-arrestins to regulate subsequent desensitization, biased signaling and endocytosis of the GPCR. The outcome of agonist-internalized GPCR in endosomes is also regulated by sequence motifs or “barcodes” within the GPCR that mediate its recycling to the plasma membrane or retention and eventual degradation as well as its subsequent signaling in endosomes. Given the vast number of diverse sequences in GPCR, several trafficking mechanisms for endosomal GPCR have been described. The majority of recycling GPCR, are sorted out of endosomes in a “sequence-dependent pathway” anchored around a type-1 PDZ-binding module found in their C-tails. For a subset of these GPCR, a second “barcode” imprinted onto specific GPCR serine/threonine residues by compartmentalized kinase networks was required for their efficient recycling through the “sequence-dependent pathway”. Mutating the serine/threonine residues involved, produced dramatic effects on GPCR trafficking, indicating that they played a major role in setting the trafficking itinerary of these GPCR. While endosomal SNX27, retromer/WASH complexes and actin were required for efficient sorting and budding of all these GPCR, additional proteins were required for GPCR sorting via the second “barcode”. Here we will review recent developments in GPCR trafficking in general and the human ß1-adrenergic receptor in particular across the various trafficking roadmaps. In addition, we will discuss the role of GPCR trafficking in regulating endosomal GPCR signaling, which promote biochemical and physiological effects that are distinct from those generated by the GPCR signal transduction pathway in membranes.

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Distinct phosphorylation sites on the ghrelin receptor, GHSR1a, establish a code that determines the functions of ß-arrestins

Cited by 40 publications

References 33 publications

Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Updates in the function and regulation of α₁‐adrenoceptors

Barcoding of GPCR trafficking and signaling through the various trafficking roadmaps by compartmentalized signaling networks

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Distinct phosphorylation sites on the ghrelin receptor, GHSR1a, establish a code that determines the functions of ß-arrestins

Cited by 40 publications

References 33 publications

Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Identification of key phosphorylation sites in PTH1R that determine arrestin3 binding and fine-tune receptor signaling

Updates in the function and regulation of α1‐adrenoceptors

Barcoding of GPCR trafficking and signaling through the various trafficking roadmaps by compartmentalized signaling networks

Contact Info

Product

Resources

About

Updates in the function and regulation of α₁‐adrenoceptors