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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 GPR150 Receptor in transfected HEK293 cells.
GPR150 (non-phospho), G protein-coupled...
The non-phospho-GPR150 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR150. It can be used to detect total GPR150 receptors in Western blots independent of phosphorylation. The GPR150...
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Validation of the GPR142 Receptor in transfected HEK293 cells.
GPR142 (non-phospho), G protein-coupled...
The non-phospho-GPR142 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR142. It can be used to detect total GPR142 receptors in Western blots independent of phosphorylation. The GPR142...
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Validation of the GPR135 Receptor in transfected HEK293 cells.
GPR135 (non-phospho), G protein-coupled...
The non-phospho-GPR135 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR135. It can be used to detect total GPR135 receptors in Western blots independent of phosphorylation. The GPR135...
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Validation of the GPR85 Receptor in transfected HEK293 cells.
GPR85 (non-phospho), G protein-coupled Receptor...
The non-phospho-GPR85 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR85. It can be used to detect total GPR85 receptors in Western blots independent of phosphorylation. The GPR85 antibody...
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Validation of the GPR68 Receptor in transfected HEK293 cells.
GPR68 (non-phospho), G protein-coupled Receptor...
The non-phospho-GPR68 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR68. It can be used to detect total GPR68 receptors in Western blots independent of phosphorylation. The GPR68 antibody...
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Validation of the GPR45 Receptor in transfected HEK293 cells.
GPR45 (non-phospho), G protein-coupled Receptor...
The non-phospho-GPR45 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR45. It can be used to detect total GPR45 receptors in Western blots independent of phosphorylation. The GPR45 antibody...
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Validation of the GPR37L1 Receptor in transfected HEK293 cells
GPR37L1 (non-phospho), G protein-coupled...
The non-phospho-GPR37L1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR37L1. It can be used to detect total GPR37L1 receptors in Western blots independent of phosphorylation. The GPR37L1...
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Validation of the GPR37 Receptor in transfected HEK293 cells
GPR37 (non-phospho), G protein-coupled Receptor...
The non-phospho-GPR37 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR37. It can be used to detect total GPR37 receptors in Western blots independent of phosphorylation. The GPR37 antibody...
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Validation of the BLT1 Receptor in transfected HEK293 cells.
BLT1 (non-phospho), Leukotriene Receptor BLT1...
The non-phospho-BLT1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human BLT1. It can be used to detect total BLT1 receptors in Western blots independent of phosphorylation. The BLT1 antibody can...
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Detection of non-phosphorylated human FFA4 Receptor
FFA4 (non-phospho-FFA4 Antibody)
The non-phospho-FFA4 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human FFA4. It can be used to detect total FFA4 receptors in Western blots independent of phosphorylation. The non-phospho-FFA4...
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Validation of the FFA2 in transfected HEK293 cells
FFA2 (non-phospho-FFA2 Antibody)
The non-phospho-FFA2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human FFA2. It can be used to detect total FFA2 receptors in Western blots independent of phosphorylation. The non-phospho-FFA2...
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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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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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