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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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Validation of the Orexin Receptor 2 in transfected HEK293 cells
OX2 (non-phospho) Orexin Receptor 2 Antibody
The non-phospho OX2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human OX2 receptor. It can be used to detect total OX2 receptors in Western blots independent of phosphorylation. It can also be used...
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Validation of the TP Tromboxane Receptor in transfected HEK293 cells
TP (non-phospho), TP Thromboxane Receptor Antibody
The non-phospho-TP Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human TP. It can be used to detect total TP receptors in Western blots independent of phosphorylation. The non-phospho-TP antibody...
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Validation of the IP Prostacyclin Receptor in transfected HEK293 cells
IP (non-phospho), IP Prostacyclin Receptor...
The non-phospho-IP Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human IP. It can be used to detect total IP receptors in Western blots independent of phosphorylation. The non-phospho-IP antibody...
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Validation of the FP Prostanoid Receptor in transfected HEK293 cells
FP (non-phospho), FP Prostanoid Receptor Antibody
The non-phospho-FP Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human FP. It can be used to detect total FP receptors in Western blots independent of phosphorylation. The non-phospho-FP antibody...
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Validation of the Hydroxycarboxylic Acid Receptor 2 in transfected HEK293 cells
HCA2 (GP-non-phospho) Hydroxycarboxylic Acid...
The non-phospho HCA2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human HCA2 receptor. It can be used to detect total HCA2 receptors in Western blots independent of phosphorylation. It can also be...
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Immunohistochemical identification of SUCNR1 Receptor in Pancreas
SUCNR1/GPR91 (IHC-grade), Succinate Receptor...
The SUCNR1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human SUCNR1. It can be used to detect total SUCNR1 receptors in Western blots independent of phosphorylation. The SUCNR1 antibody can also...
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Agonist-induced Serine369/Serine370 phosphorylation of the EP3 Prostanoid Receptor
pS369/pS370-EP3 (phospho-EP3 Prostanoid...
Serine369/Serine370 (S369/S370) is major phosphorylation site of the EP3 Prostanoid Receptor (EP3). The pS369/pS370-EP3 antibody detects phosphorylation in response to agonists. S369/S370 phosphorylation is likely to be involved in...
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Agonist-induced Threonine372/Serine373 phosphorylation of the EP3 Prostanoid Receptor
pT372/pS373-EP3 (phospho-EP3 Prostanoid...
Threonine372/Serine373 (T372/S373) is major phosphorylation site of the EP3 Prostanoid Receptor (EP3). The pT372/pS373-EP3 antibody detects phosphorylation in response to agonists. T372/S373 phosphorylation is likely to be involved in...
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Agonist-induced Serine379/Serine380 phosphorylation of the EP3 Prostanoid Receptor
pS379/pS380-EP3 (phospho-EP3 Prostanoid...
Serine379/Serine380 (S379/S380) is major phosphorylation site of the EP3 Prostanoid Receptor (EP3). The pS379/pS380-EP3 antibody detects phosphorylation in response to agonists. S379/S380 phosphorylation is likely to be involved in...
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pS338/pS339-DP2 (phospho-DP2 Prostanoid Receptor Antibody)
pS338/pS339-DP2 (phospho-DP2 Prostanoid...
Serine338/Serine339 (S338/S339) is major phosphorylation site of the DP2 Prostanoid Receptor (DP2). The pS338/pS339-DP2 antibody detects phosphorylation in response to agonists. S338/S339 phosphorylation is likely to be involved in...
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Agonist-induced Threonine344/Serine345 phosphorylation of the Prostanoid Receptor 2
pT344/pS345-DP2 (phospho-DP2 Prostanoid...
Threonine344/Serine345 (T344/S345) is major phosphorylation site of the DP2 Prostanoid Receptor (DP2). The pT344/pS345-DP2 antibody detects phosphorylation in response to agonists. T344/S345 phosphorylation is likely to be involved in...
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Agonist-induced Serine350 phosphorylation of the Vasopressin Receptor 2
pS350-V2 (phospho-Vasopressin Receptor 2 Antibody)
Serine350 (S350) is major phosphorylation site of the Vasopresssin Receptor 2 (V2). The pS350-V2 antibody detects phosphorylation in response to agonists. S350 phosphorylation is likely to be involved in efficient ligand sequestration by...
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Agonist-induced Threonine359/Threonine360 phosphorylation of the Vasopressin Receptor 2.
pT359/pT360-V2 (phospho-Vasopressin Receptor 2...
Threonine359/Threonine360 (T359/T360) is major phosphorylation site of the Vasopressin Receptor 2 (V2). The pT359/pT360-V2 antibody detects phosphorylation in response to agonists. T359/T360 phosphorylation is likely to be involved in...
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Agonist-induced Serine362/Serine363/Serine364 phosphorylation of the Vasopressin Receptor 2
pS362/pS363/pS364-V2 (phospho-Vasopressin...
Serine362/Serine363/Serine364 (S362/S363/S364) is major phosphorylation site of the Vasopresssin Receptor 2 (V2). The pS362/pS363/pS364-V2 antibody detects phosphorylation in response to agonists. S362/S363/S364 phosphorylation is likely...
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pS373/pS374-V1B (phospho-Vasopressin Receptor 1B Antibody)
pS373/pS374-V1B (phospho-Vasopressin Receptor...
Serine373/Serine374 (S373/S374) is major phosphorylation site of the Vasopressin Receptor 1B (V1B). The pS373/pS374-V1B antibody detects phosphorylation in response to agonists. S373/S374 phosphorylation is likely to be involved in...
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Agonist-induced Threonine377/Threonine378 phosphorylation of the Vasopressin Receptor 1B
pT377/pT378-V1B (phospho-Vasopressin Receptor...
Threonine377/Threonine378 (T377/T378) is major phosphorylation site of the Vasopressin Receptor 1B (V1B). The pT377/pT378-V1B antibody detects phosphorylation in response to agonists. T377/T378 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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