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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 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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Validation of the GPR176 Receptor in transfected HEK293 cells.
GPR176 (non-phospho), G protein-coupled...
The non-phospho-GPR176 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR176. It can be used to detect total GPR176 receptors in Western blots independent of phosphorylation. The GPR176...
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Validation of the APLN Receptor in transfected HEK293 cells
APLNR (non-phospho), Apelin Receptor Antibody
The non-phospho-APLNR antibody is directed against the distal end of the carboxyl-terminal tail of human apelin receptor. It also detects APLNR in cultured cells and tissue sections by immunohistochemistry. It can be used to detect total...
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Validation of the S1P1 Receptor in transfected HEK293 cells
S1P1 (non-phospho), Sphingosine 1-Phosphate...
The non-phospho-S1P1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human S1P1. It also detects S1P1 in cultured cells and tissue sections by immunohistochemistry. It can be used to detect total...
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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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Agonist-induced Serine339/Serine340 phosphorylation of GAL2 Receptor
pS339/pS340-GAL2 (phospho-Galanin Receptor 2...
Serine339/Serine340 (S339/S340) is a major phosphorylation site of the GAL2 receptor. The pS339/pS340-GAL2 antibody detects phosphorylation in response to high-efficacy agonists. S339/S340 phosphorylation is a key regulator of GAL2...
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Agonist-induced Serine331/Serine333 phosphorylation of GAL2 Receptor.
pS331/pS333-GAL2 (phospho-Galanin Receptor 2...
Serine331/Serine333 (S331/S333) is a major phosphorylation site of the GAL2 receptor. The pS331/pS333-GAL2 antibody detects phosphorylation in response to high-efficacy agonists. S331/S333 phosphorylation is a key regulator of GAL2...
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Agonist-induced Threonine329/Serine331 phosphorylation of GAL2 Receptor
pT329/pS331-GAL2 (phospho-Galanin Receptor 2...
Threonine329/Serine331 (T329/S331) is a major phosphorylation site of the GAL2 receptor. The pT329/pS331-GAL2 antibody detects phosphorylation in response to high-efficacy agonists. T329/S331 phosphorylation is a key regulator of GAL2...
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Agonist-induced Threonine353/Threonine356 phosphorylation of NK2 Receptor.
pT353/pT356-NK2 (phospho-Tachykinin Receptor 2...
Threonine353/Threonine356 (T353/T356) is a major phosphorylation site of the NK2 receptor. The pT353/pT356-NK2 antibody detects phosphorylation in response to high-efficacy agonists. T353/T356 phosphorylation is a key regulator of NK2...
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Agonist-induced Serine345/Threonine346 phosphorylation of NK2 Receptor.
pS345/pT346-NK2 (phospho-Tachykinin Receptor 2...
Serine345/Threonine346 (S345/T346) is a major phosphorylation site of the NK2 receptor. The pS345/pT346-NK2 antibody detects phosphorylation in response to high-efficacy agonists. S345/T346 phosphorylation is a key regulator of NK2...
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Agonist-induced Threonine339/Threonine341 phosphorylation of NK2 Receptor.
pT339/pT341-NK2 (phospho-Tachykinin Receptor 2...
Threonine339/Threonine341 (T339/T341) is a major phosphorylation site of the NK2 receptor. The pT339/pT341-NK2 antibody detects phosphorylation in response to high-efficacy agonists. T339/T341 phosphorylation is a key regulator of NK2...
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Serine358/Seine362 phosphorylation of the G-protein coupled Receptor 176.
pS358/pS362-GPR176 (phospho-GPR176 Antibody)
Serine358/Serine362 (S358/S362) is a major phosphorylation site of the human GPR176 receptor. The pS358/pS362-GPR176 antibody detects GRK-mediated constitutive phosphorylation as well as PKC-mediated phosphorylation of GPR176 when...
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Threonine427/Serine429 phosphorylation of the G-protein coupled Receptor 75.
pT427/pS429-GPR75 (phospho-GPR75 Antibody)
Threonine427/Serine429 (T427/S429) is a major phosphorylation site of the human GPR75 receptor. The pT427/pS429-GPR75 antibody detects GRK-mediated constitutive phosphorylation of GPR75 when expressed in HEK293 cells.
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Serine422/Serine423 phosphorylation of the G-protein coupled Receptor 75
pS422/pS423-GPR75 (phospho-GPR75 Antibody)
Serine422/Serine423 (S422/S423) is a major phosphorylation site of the human GPR75 receptor. The pS422/pS423-GPR75 antibody detects GRK-mediated constitutive phosphorylation as well as PKC-mediated phosphorylation of GPR75 when expressed...
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Validation of the Sphingosine 1-Phosphate Receptor 3 in transfected HEK293 cells.
S1P3 (non-phospho), Sphingosine 1-Phosphate...
The non-phospho-sphingosin 1-phosphate receptor 3 antibody is directed against the distal end of the carboxyl-terminal tail of human S1P3.
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Agonist-induced Threonine408/Threonine409 phosphorylation of Orexin Receptor 2
pT408/pT409-OX2 (phospho-Orexin Receptor 2...
Threonine408/Threonine409 (T408/T409) is a major phosphorylation site of the Orexin receptor 2 (OX2). The pT408/pT409-OX2 antibody detects phosphorylation in response to high-efficacy agonists. T408/T409 phosphorylation is a key...
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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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Premium Phosphosite-Specific 7TM Antibodies

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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