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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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Agonist-induced Threonine378/Serine380 phosphorylation of the Vasopressin Receptor 1A
pT378/pS380-V1A (phospho-Vasopressin Receptor...
Threonine378/Serine380 (T378/S380) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pT378/pS380-V1A antibody detects phosphorylation in response to agonists. T378/S380 phosphorylation is likely to be involved in...
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Agonist-induced Threonine386/Serine389 phosphorylation of the Vasopressin Receptor 1A
pT386/pS389-V1A (phospho-Vasopressin Receptor...
Threonine386/Serine389 (T368/S389) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pT386/pS389-V1A antibody detects phosphorylation in response to agonists. T386/S389 phosphorylation is likely to be involved in...
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Agonist-induced Serine397/Threonine398 phosphorylation of the Vasopressin Receptor 1A
pS397/pT398-V1A (phospho-Vasopressin Receptor...
Serine397/Threonine398 (S397/T398) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pS397/pT398-V1A antibody detects phosphorylation in response to agonists. S397/T398 phosphorylation is likely to be involved in...
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Agonist-induced Serine339 phosphorylation of the P2Y Receptor 4
pS339-P2Y4 (phospho-P2Y Purinoceptor 4 Antibody)
Serine339 (S339) is a major phosphorylation site of the P2Y4 receptor. The pS339-P2Y4 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S339 phosphorylation is a key regulator of...
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Agonist-induced Threonine351 phosphorylation of the P2Y Receptor 4
pT351-P2Y4 (phospho-P2Y Purinoceptor 4 Antibody)
Threonine351 (T351) is a major phosphorylation site of the P2Y4 receptor. The pT351-P2Y4 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T351 phosphorylation is a key regulator of...
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Agonist-induced Serine366/Serine368 phosphorylation of the Oxytocin Receptor
pS366/pS368-OT (phospho-Oxytocin Receptor...
Serine366/Serine368 (S366/S368) is major phosphorylation site of the Oxytocin Receptor (OT). The pS366/pS368-OT antibody detects phosphorylation in response to agonists. S366/S368 phosphorylation is likely to be involved in efficient...
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Agonist-induced Serine377/Ser378 phosphorylation of the Oxytocin Receptor.
pS377/pS378-OT (phospho-Oxytocin Receptor...
Serine377/Serine378 (S377/S378) is major phosphorylation site of the Oxytocin Receptor (OT). The p3S77/pS378-OT antibody detects phosphorylation in response to agonists. S377/S378 phosphorylation is likely to be involved in efficient...
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Agonist-induced Serine361 phosphorylation of the Neuropeptide FF Receptor 1
pS361-NPFF1 (phospho-Neuropeptide FF Receptor 1...
Serine361 (S361) is major phosphorylation site of the Neuropeptide FF Receptor 1 (NPFF1). The pS361-NPFF1 antibody detects phosphorylation in response to agonists. S361 phosphorylation is likely to be involved in efficient ligand...
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Agonist-induced Serine385/Serine387 phosphorylation of the Neuropeptide Receptor 1
pS385/pS387-NPFF1 (phospho-Neuropeptide FF...
Serine385/Serine387 (S385/S387) is major phosphorylation site of the Neuropeptide FF Receptor 1 (NPFF1). The pS385/pS387-NPFF1 antibody detects phosphorylation in response to agonists. S385/S387 phosphorylation is likely to be involved...
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Agonist-induced Threonine389/Threonine390 phosphorylation of the NK3 Receptor3.
pT389/pT390-NK3 (phospho-Tachykinin Receptor 3...
Threonine389/Threonine390 (T389/T390) is a major phosphorylation site of the NK3 receptor. The pT389/pT390-NK3 antibody detects phosphorylation in response to high-efficacy agonists. T389/T390 phosphorylation is a key regulator of NK3...
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Agonist-induced Serine398/Serine399 phosphorylation of the NK 3 Recept
pS398/pS399-NK3 (phospho-Tachykinin Receptor 3...
Serine398/Serine399 (S398/S399) is major phosphorylation site of the Tachykinin Receptor 3 (NK3). The pS398/pS399-NK3 antibody detects phosphorylation in response to agonists. S398/S399 phosphorylation is likely to be involved in...
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Agonist-induced Threonine402/Threonine404 phosphorylation of the NK 3 Receptor.
pT402/pT404-NK3 (phospho-Tachykinin Receptor 3...
Threonine402/Threonine404 (T402/T404) is a major phosphorylation site of the NK3 receptor. The pT402/pT404-NK3 antibody detects phosphorylation in response to high-efficacy agonists. T402/T404 phosphorylation is a key regulator of NK3...
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Agonist-induced Serine313 phosphorylation of the MRGPRX2 Mas-related Receptor
pS313-MRGPRX2 (phospho-MRGPRX2 Mas-related...
Serine313 (S313) is a major phosphorylation site of the MRGPRX2 Mas-related receptor. The pS313-MRGPRX2 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S313 phosphorylation is a...
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Agonist-induced Serine327/Serine328 phosphorylation of the MRGPRX2 Mas-related Receptor
pS327/pS328-MRGPRX2 (phospho-MRGPRX2...
Serine327/Serine328 (S327/S329) is a major phosphorylation site of the MRGPRX2 Mas-related receptor. The pS327/pS328-MRGPRX2 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation....
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Immunohistochemical identification of the Formylpeptide Receptor 2 in striatum
FPR2 (IHC-grade), Formylpeptide Receptor 2...
The FPR2 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human FPR2. It can be used to detect total FPR2 receptors in Western blots independent of phosphorylation. The FPR2 antibody can also be...
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Agonist-induced Threonine337/Serine340 phosphorylation of the TP Thromboxane Receptor
pT337/pS340-TP (phospho-TP Thromboxane Receptor...
Threonine337/Serine340 (T337/S340) is major phosphorylation site of the TP Thromboxane Receptor (TP). The pT337/pS340-TP antibody detects phosphorylation in response to agonists. T337/S340 phosphorylation is likely to be involved in...
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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.

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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