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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 M2 Muscarinic Acetylcholine Receptor in transfected HEK293 cells
M2 (non-phospho-M2 Muscarinic Acetylcholine...
The non-phospho-M2 receptor antibody is directed against the 3rd loop of human M2 Muscarinic Acetylcholine Receptor (M2). It can be used to detect total M2 receptors in Western blots independent of phosphorylation. The non-phospho-M2...
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Validation of the EP2 Receptor in transfected HEK293 cells
EP2 (non-phospho), EP2 Prostanoid Receptor...
The non-phospho-EP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human EP2. It can be used to detect total EP2 receptors in Western blots independent of phosphorylation.The EP2 antibody can also...
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Agonist-induced Serine377 phosphorylation of QRFP Receptor
pS377-QRFPR (phospho-QRFP Receptor Antibody)
Serine377 (S377) is a major phosphorylation site of the QRFP receptor. The pS377-QRFPR antibody detects phosphorylation in response to high-efficacy agonists. S377 phosphorylation is a key regulator of QRFPR desensitization, β-arrestin...
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Agonist-induced Serine365/Threonine368 phosphorylation of QRFP Receptor
pS365/pT368-QRFPR (phospho-QRFP Receptor Antibody)
Serine365/Threonine368 (S365/T368) is a major phosphorylation site of the QRFP receptor. The pS365/pT368-QRFPR antibody detects phosphorylation in response to high-efficacy agonists. S365/T368 phosphorylation is a key regulator of QRFPR...
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Agonist-induced Threonine348/Serine350 phosphorylation of MCH1 Receptor
pT348/pS350-MCH1 (phospho-Melanin-Concentrating...
Threonine348/Serine350 (T348/S350) is a major phosphorylation site of the MCH1 receptor. The pT348/pS350-MCH1 antibody detects phosphorylation in response to high-efficacy agonists. T348/S350 phosphorylation is a key regulator of MCH1...
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Agonist-induced Serine338/Threonine342 phosphorylation of MCH1 Receptor.
pS338/pT342-MCH1 (phospho-Melanin-Concentrating...
Serine338/Threonine342 (S338/T342) is a major phosphorylation site of the MCH1 receptor. The pS338/pT342-MCH1 antibody detects phosphorylation in response to high-efficacy agonists. S338/T342 phosphorylation is a key regulator of MCH1...
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Validation of the GPR158/mGlyR Receptor in transfected HEK293 cells
GPR158/mGlyR (non-phospho), G protein-coupled...
The non-phospho-GPR158/mGlyR receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR158/mGlyR. It can be used to detect total GPR158/mGlyR receptors in Western blots independent of phosphorylation....
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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 GPR75 Receptor in transfected HEK293 cells.
GPR75 (non-phospho), G protein-coupled...
The non-phospho-GPR75 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR75. It can be used to detect total GPR75 receptors in Western blots independent of phosphorylation. The GPR75 antibody...
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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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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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