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Phosphorylation of intracellular serine and threonine residues is the most important post translational modification of G protein-coupled receptors (GPCRs) also called heptahelical or seven transmembrane receptors (7TMR). After agonist exposure, these receptors acquire an active conformation, which is recognized by a family of highly specialized GPCR kinases (GRKs). Agonist-driven phosphorylation by GRKs regulates acute receptor desensitization, arrestin recruitment, internalization, post-activation signaling, long-term tolerance and drug addiction. Phosphosite-specific 7TM antibodies are designed to specifically detect agonist-activated GPCRs. In fact, recent work shows that ligand profiling using phosphosite-specific 7TM antibodies provides valuble information on ligand bias beyond that obtained with conventional ß-arrestin recruitment assays. Phosphosite-specific 7TM antibodies are novel tools for GPCR research that can be used to:

  • profile agonist properties of novel GPCR ligands
  • decipher the phosphorylation barcode of GPCRs
  • determine the spatial and temporal dynamics of receptor phosphorylation
  • identify relevant kinases and phosphatases for GPCR phosphorylation and dephosphoryation

Lifecycle3

Schematic representation of the G protein-coupled receptor phosphorylation / dephosphorylation cycle. GRK, G protein-coupled receptor kinase; PKC, protein kinase C; cPP1, catalytic subunit of protein phosphatase 1; R*, activated GPCR; CCP, clathrin-coated pit. 

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Immunohistochemical identification of G Protein-coupled Receptor 82 in hippocampus
GPR82 (IHC-grade), G Protein-Coupled Receptor...
The GPR82 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP82. It can be used to detect total GPR82 receptors in Western blots independent of phosphorylation. The GPR82 antibody can...
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Immunohistochemical identification of G Protein-coupled Receptor 65 in small intestine
GPR65 (IHC-grade), G Protein-Coupled Receptor...
The GPR65 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP65. It can be used to detect total GPR65 receptors in Western blots independent of phosphorylation. The GPR65 antibody can...
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Immunohistochemical identification of G Protein-coupled Receptor 61 in cortex
GPR61 (IHC-grade), G Protein-Coupled Receptor...
The GPR61 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP61. It can be used to detect total GPR61 receptors in Western blots independent of phosphorylation. The GPR61 antibody can...
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Immunohistochemical identification of GPR3 Receptor in Cortex
GPR3 (IHC-grade), G Protein-Coupled Receptor 3...
The GPR3 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP3. It can be used to detect total GPR3 receptors in Western blots independent of phosphorylation. The GPR3 antibody can also be...
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Agonist-induced Serine331/Threonine332 phosphorylation of the Lysophosphatidic Acid Receptor 2
pS331/pT332-LPA2 (phospho-Lysophosphatidic Acid...
Serine331/Threonine332 (S331/T332) is major phosphorylation site of the Lysophosphatidic Acid Receptor 2 (LPA2). pS331/pT332-LPA2 antibody detects phosphorylation in response to agonists. S331/T332 phosphorylation is likely to be...
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Agonist-induced Threonine323/Serine324/Serine325 phosphorylation of the Lysophosphatidic Acid Receptor 2
pT323/pS324/pS325-LPA2...
Threonine323/Serine324/Serine325 (T323/S324/325) is major phosphorylation site of the Lysophosphatidic Acid Receptor 2 (LPA2). pT323/pS324/pS325-LPA2 antibody detects phosphorylation in response to agonists. T323/S324/325 phosphorylation...
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Agonist-induced Threonine332/Serine335 phosphorylation of the Lysophosphatidic Acid Receptor 6
pT332/pS335-LPA6 (phospho-Lysophosphatidic Acid...
Threonine332/Serine335 (T332/S335) is major phosphorylation site of the Lysophosphatidic Acid Receptor 6 (LPA6). pT332/pS335-LPA6 antibody detects phosphorylation in response to agonists. T332/S335 phosphorylation is likely to be...
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Agonist-induced Threonine338/Serine341 phosphorylation of the Lysophosphatidic Acid Receptor 1
pT338/pS341-LPA1 (phospho-Lysophosphatidic Acid...
Threonine338/Serine341 (T338/S341) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pT338/pS341-LPA1 antibody detects phosphorylation in response to agonists. T338/S341 phosphorylation is likely to be...
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Agonist-induced Threonine336/Threonine338 phosphorylation of the Lysophosphatidic Acid Receptor 4
pT336/pT338-LPA4 (phospho-Lysophosphatidic Acid...
Threonine336/Threonine338 (T336/T338) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pT336/pT338-LPA4 antibody detects phosphorylation in response to agonists. T336/T338 phosphorylation is likely to be...
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Agonist-induced Serine354/Threonine357/Threonine358 phosphorylation of the Lysophosphatidic Acid Receptor 4
pS354/pT357/pT358-LPA4...
Serine354/Threonine357/Threonine358 (S354/T357/T358) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pS354/pT357/pT358-LPA4 antibody detects phosphorylation in response to agonists. S354/T357/T358...
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Agonist-induced Threonine341/Threonine342/Serine345 phosphorylation of the Lysophosphatidic Acid Receptor 4
pT341/pT342/pS345-LPA4...
Threonine341/Threonine342/Serine345 (T341/T342/S345) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pT341/pT342/pS345-LPA4 antibody detects phosphorylation in response to agonists. T341/T342/S345...
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Agonist-induced Serine354/Serine359 phosphorylation of the Relaxin Receptor 4
pS354/pS359-RXFP4 (phospho-Relaxin Receptor 4...
Serine354/Serine359 (S354/S359) is major phosphorylation site of the Relaxin Receptor 4 (RXFP4). The pS354/pS359-RXFP4 antibody detects phosphorylation in response to agonists. S354/S359 phosphorylation is likely to be involved in...
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Agonist-induced Threonine419/Threonine421/Threonine422 phosphorylation of the Relaxin Receptor 3
pT419/pT421/pT422-RXFP3 (phospho-Relaxin...
Threonine419/Threonine421/Threonine422 (T419/T421/T422) is major phosphorylation site of the Relaxin Receptor 3 (RXFP3). The pT419/pT421/pT422-RXFP3 antibody detects phosphorylation in response to agonists. T419/T421/T422 phosphorylation...
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Agonist-induced Serine368 phosphorylation of the Kisspeptin Receptor
pS368-KISS1R (phospho-Kisspeptin Receptor...
Serine368 (S368) is major phosphorylation site of the Kisspeptin Receptor (KISS1R). The pS368-KISS1R antibody detects phosphorylation in response to agonists. S368 phosphorylation is likely to be involved in efficient ligand...
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Agonist-induced Threonine351 phosphorylation of the Lysophosphatidic Acid Receptor 1
pT351-LPA1 (phospho-Lysophosphatidic Acid...
Threonine351 (T351) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pT351-LPA1 antibody detects phosphorylation in response to agonists. T351 phosphorylation is likely to be involved in efficient ligand...
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Agonist-induced Serine344/Serine346/Serine347 phosphorylation of the Lysophosphatidic Acid Receptor 1
pS344/pS346/pS347-LPA1...
Serine344/Serine346/Serine347 (S344/S346/S347) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pS344/pS346/pS347-LPA1 antibody detects phosphorylation in response to agonists. S344/S346/S347...
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11 From 37

For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

For further reading refer to: Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636.... read more »
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For further reading refer to:

Kliewer A, Reinscheid RK, Schulz S. Emerging Paradigms of G Protein-Coupled Receptor Dephosphorylation. Trends Pharmacol Sci. 2017 Jul;38(7):621-636. doi:10.1016/j.tips.2017.04.002. Epub 2017 May 4. Review. PubMed PMID: 28478994.

Miess E, Gondin AB, Yousuf A, Steinborn R, Mösslein N, Yang Y, Göldner M, Ruland JG, Bünemann M, Krasel C, Christie MJ, Halls ML, Schulz S, Canals M. Multisite phosphorylation is required for sustained interaction with GRKs and arrestins during rapid μ-opioid receptor desensitization. Sci Signal. 2018 Jul 17;11(539). pii: eaas9609. doi: 10.1126/scisignal.aas9609. PubMed PMID: 30018083.

Kliewer A, Schmiedel F, Sianati S, Bailey A, Bateman JT, Levitt ES, Williams JT, Christie MJ, Schulz S. Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia and diminish tolerance but worsen opioid side effects. Nat Commun. 2019 Jan 21;10(1):367. doi: 10.1038/s41467-018-08162-1. PubMed PMID: 30664663; PubMed Central PMCID: PMC6341117.

Mann A, Moulédous L, Froment C, O'Neill PR, Dasgupta P, Günther T, Brunori G, Kieffer BL, Toll L, Bruchas MR, Zaveri NT, Schulz S. Agonist-selective NOP receptor phosphorylation correlates in vitro and in vivo and reveals differential post-activation signaling by chemically diverse agonists. Sci Signal. 2019 Mar 26;12(574). pii: eaau8072. doi: 10.1126/scisignal.aau8072. PubMed PMID: 30914485; PubMed Central PMCID: PMC6934085.

Saaber F, Schütz D, Miess E, Abe P, Desikan S, Ashok Kumar P, Balk S, Huang K, Beaulieu JM, Schulz S, Stumm R. ACKR3 Regulation of Neuronal Migration Requires ACKR3 Phosphorylation, but Not β-Arrestin. Cell Rep. 2019 Feb 5;26(6):1473-1488.e9. doi: 10.1016/j.celrep.2019.01.049. PubMed PMID: 30726732.

Glück L, Loktev A, Moulédous L, Mollereau C, Law PY, Schulz S. Loss of morphine reward and dependence in mice lacking G protein-coupled receptor kinase 5. Biol Psychiatry. 2014 Nov 15;76(10):767-74. doi: 10.1016/j.biopsych.2014.01.021. Epub 2014 Feb 3. PubMed PMID: 24629717; PubMed Central PMCID: PMC4119866.

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