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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 Threonine344/Serine347 phosphorylation of NK1 Receptor
pT344/pS347-NK1 (phospho-Tachykinin Receptor 1...
Threonine344/Serine347 (T344/S347) is a major phosphorylation site of the NK1 receptor. The pT344/pS347-NK1 antibody detects phosphorylation in response to high-efficacy agonists. T344/S347 phosphorylation is a key regulator of NK1...
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NEW
Validation of the Proteinase-Activated Receptor 2 in transfected HEK293 cells
PAR2 (non-phospho), Proteinase-Activated...
The non-phospho-PAR2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Proteinase-Activated Receptor 2. It can be used to detect total PAR2 receptors in Western blots independent of...
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NEW
Validation of the GPR132 Receptor in transfected HEK293 cells
GPR132 (non-phospho), G protein-coupled...
The non-phospho-GPR132 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR132. It can be used to detect total GPR132 receptors in Western blots independent of phosphorylation. The GPR132...
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Agonist-induced Threonine315 phosphorylation of the GPR139 Receptor
pT315-GPR139 (phospho-GPR139 Antibody)
Threonine315 (T315) is major phosphorylation site of the GPR139 Receptor (GPR139). The pT315-GPR139 antibody detects phosphorylation in response to agonists. T315 phosphorylation is likely to be involved in efficient ligand sequestration.
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Agonist-induced Serine323/Serine324 phosphorylation of the GPR139 Receptor
pS323/pS324-GPR139 (phospho-GPR139 Antibody)
Serine323/Serine324 (S323/S324) is major phosphorylation site of the GPR139 Receptor (GPR139). The pS323/pS324-GPR139 antibody detects phosphorylation in response to agonists. S323/S324 phosphorylation is likely to be involved in...
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Validation of the mouse S1P5 Receptor in transfected HEK293 cells
mS1P5 (GP-non-phospho), Mouse Sphingosine...
The non-phospho-mS1P5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouseS1P5. It can be used to detect total mS1P5 receptors in Western blots independent of phosphorylation. The mS1P5 antibody can...
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Validation of the GPR88 Receptor in transfected HEK293 cells
GPR88 (non-phospho), G protein-coupled...
The non-phospho-GPR88 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR88. It can be used to detect total GPR88 receptors in Western blots independent of phosphorylation. The GPR88 antibody...
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Agonist-induced Serine346/Serine349 phosphorylation of GPR88-Receptor
pS346/pS349-GPR88 (phospho-GPR88 Antibody)
Serine346/Serine349 (S346/S349) is a major phosphorylation site of the human GPR88 receptor. The pS346/pS349-GPR88 antibody detects phosphorylation in response to high-efficacy agonists. S346/S349 phosphorylation is a key regulator of...
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Agonist-induced Serine371 phosphorylation of GPR88-Receptor.
pS371-GPR88 (phospho-GPR88 Antibody)
Serine371 (S371) is a major phosphorylation site of the human GPR88 receptor. The pS371-GPR88 antibody detects phosphorylation in response to high-efficacy agonists. S371 phosphorylation is a key regulator of human GPR88 desensitization,...
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Agonist-induced Threonine377 phosphorylation of GPR88-Receptor
pT377-GPR88 (phospho-GPR88 Antibody)
Threonine377 (T377) is a major phosphorylation site of the human GPR88 receptor. The pT377-GPR88 antibody detects phosphorylation in response to high-efficacy agonists. T377 phosphorylation is a key regulator of human GPR88...
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Immunohistochemical identification of Threonine344/Serine347 phosphorylation of NK1 Receptor in striatum
pT344/pS347-NK1 (IHC-grade phospho-Tachykinin...
Threonine344/Serine347 (T344/S347) is a major phosphorylation site of the NK1 receptor. The pT344/pS347-NK1 antibody detects phosphorylation in response to high-efficacy agonists. T344/S347 phosphorylation is a key regulator of NK1...
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Validation of the GPR52 Receptor in transfected HEK293 cells
GPR52 (non-phospho), G protein-coupled...
The non-phospho-GPR52 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR52. It can be used to detect total GPR52 receptors in Western blots independent of phosphorylation. The GPR52 antibody...
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Immunohistochemical varification of 5-Hydroxytryptamine Receptor 6 antibody in Medial Septum
5-HT6 (IHC-grade), 5-HT Receptor 6 Antibody
The 5-HT6 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human 5-Hydroxytryptamine Receptor 6 (5-HT6). It can be used to detect total 5-HT6 receptors in Western blots independent of...
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Immunohistochemical identification of Dopamine Receptor 1 in striatum
D1 (IHC-grade), Dopamine Receptor 1 Antibody
The D1 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human D1. It can be used to detect total D1 receptors in Western blots independent of phosphorylation. The D1 antibody can also be used to...
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Validation of the S1P5 Receptor in transfected HEK293 cells.
S1P5 (non-phospho), Sphingosine 1-Phosphate...
The non-phospho-S1P5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human S1P5. It can be used to detect total S1P5 receptors in Western blots independent of phosphorylation. The S1P5 antibody can...
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Detection of non-phosphorylated human GPR35 Receptor
GPR35 (non-phospho), G Protein-coupled Receptor...
The non-phospho-GPR35 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human GPR35. It can be used to detect total GPR35 receptors in Western blots independent of phosphorylation. The...
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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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