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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 GPR62 Receptor in transfected HEK293 cells
GPR62 (non-phospho), G protein-coupled...
The non-phospho-GPR62 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR62. It can be used to detect total GPR62 receptors in Western blots independent of phosphorylation. The GPR62 antibody...
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Validation of the M5 Muscarinic Acetylcholine Receptor in transfected HEK293 cells
M5 (non-phospho-M5 Muscarinic Acetylcholine...
The non-phospho-M5 receptor antibody is directed against the 3rd loop of human M5 Muscarinic Acetylcholine Receptor (M5). It can be used to detect total M5 receptors in Western blots independent of phosphorylation. The non-phospho-M5...
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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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Agonist-induced Threonine428/Serine429 phosphorylation of CRF1 Receptor
pT428/pS429-CRF1...
Threonine428/Serine429 (T428/S429) is a major phosphorylation site of the CRF1 receptor. The pT428/pS429-CRF1 antibody detects phosphorylation in response to high-efficacy agonists. T428/S429 phosphorylation is a key regulator of CRF1...
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Agonist-induced Threonine431 phosphorylation of CRF1 Receptor
pT431-CRF1 (phospho-Corticotropin-Releasing...
Threonine431 (T431) is a major phosphorylation site of the CRF1 receptor. The pT431-CRF1 antibody detects phosphorylation in response to high-efficacy agonists. T431 phosphorylation is a key regulator of CRF1 desensitization, β-arrestin...
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Agonist-induced Serine434/Serine437 phosphorylation of CRF1 Receptor
pS434/pS437-CRF1...
Serine434/Serine441 (S437/S441) is a major phosphorylation site of the CRF1 receptor. The pS434/pS437-CRF1 antibody detects phosphorylation in response to high-efficacy agonists. S434/S437 phosphorylation is a key regulator of CRF1...
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Validation of the Bombesin Receptor 2 in transfected HEK293 cells
BB2 (non-phospho), Bombesin Receptor 2 Antibody
The BB2/GRPR receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Bombesin Receptor 2 (previously called Gastrin-Releasing Peptide Receptor). It can be used to detect total BB2 receptors in Western...
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Agonist-induced Threonine340 phosphorylation of GAL1 Receptor.
pT340-GAL1 (phospho-Galanin Receptor 1 Antibody)
Threonine340 (T340) is a major phosphorylation site of the GAL1 receptor. The pT340-GAL1 antibody detects phosphorylation in response to high-efficacy agonists. T340 phosphorylation is a key regulator of GAL1 desensitization, β-arrestin...
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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 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 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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Validation of the GPR97/ADGR3G Receptor in transfected HEK293 cells
ADGRG3/GPR97 (non-phospho), Adhesion Receptor...
The non-phospho-ADGRG3/GPR97 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human ADGRG3/GPR97. It can be used to detect total ADGRG3/GPR97 receptors in Western blots independent of phosphorylation....
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Validation of the Parathyroid Hormone Receptor 1 in transfected HEK293 cells
PTH1 (non-phospho-Parathyroid Hormone Receptor...
The PTH1 antibody is directed against the distal end of the carboxyl-terminal tail of human Parathyroid Hormone Receptor 1. It can be used to detect total PTH1 receptors in Western blots independent of phosphorylation. The PTH1 antibody...
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Validation of the BLT2 Receptor in transfected HEK293 cells.
BLT2 (non-phospho), Leukotriene Receptor BLT2...
The non-phospho-BLT2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human BLT2. It can be used to detect total BLT2 receptors in Western blots independent of phosphorylation. The BLT2 antibody can...
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Agonist-induced Serine300/Serine303 phosphorylation of GPR35 Receptor.
pS300/pS303-hGPR35 (phospho-human GPR35 Antibody)
Serine300/Serine303 (S300/S303) is a major phosphorylation site of the human GPR35 receptor. The pS300/pS303-hGPR35 antibody detects phosphorylation in response to high-efficacy agonists. S300/S303 phosphorylation is a key regulator of...
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Agonist-induced Serine298/Serine301 phosphorylation of mouse GPR35 Receptor
pS298/pS301-mGPR35 (phospho-mouse GPR35 Antibody)
Serine298/Serine301 (S298/S301) is a major phosphorylation site of the mouse GPR35 receptor. The pS298/pS301-mGPR35 antibody detects phosphorylation in response to high-efficacy agonists. S298/S301 phosphorylation is a key regulator of...
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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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