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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 Serine473/Serine475 phosphorylation of the β1-Adrenoceptor
pS473/pS475-β1 (phospho-β1-Adrenoceptor Antibody)
Serine473/Serine475 (S473/S475) is a major phosphorylation site of the β1 adrenoceptor. The pS473/pS475-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S473/S475...
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Agonist-induced Serine412 phosphorylation of the β1-Adrenoceptor
pS412-β1 (phospho-β1-Adrenoceptor Antibody)
Serine412 (S412) is a major phosphorylation site of the β1 adrenoceptor. The pS412-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S412 phosphorylation is primarily...
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Agonist-induced Threonine314/Threonine315 phosphorylation of GPR55
pT314/pT315-GPR55 (phospho-GPR55 Antibody)
Threonine314/Threonine315 (T314/T315) is major phosphorylation site of human GPR55 receptor. The pT314/pT315-GPR55 antibody detects phosphorylation in response to agonists. T314/T315) phosphorylation is likely to be involved in efficient...
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Agonist-induced Threonine351 phosphorylation of the Lysophosphatidic Acid Receptor 1
pT351-LPA1 (phospho-Lysophosphatidic Acid...
Threonine351 (T351) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pT351-LPA1 antibody detects phosphorylation in response to agonists. T351 phosphorylation is likely to be involved in efficient ligand...
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Agonist-induced Serine344/Serine346/Serine347 phosphorylation of the Lysophosphatidic Acid Receptor 1
pS344/pS346/pS347-LPA1...
Serine344/Serine346/Serine347 (S344/S346/S347) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pS344/pS346/pS347-LPA1 antibody detects phosphorylation in response to agonists. S344/S346/S347...
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Agonist-induced Threoine347/Serine349 phosphorylation of GPR183
pT347/pT349-GPR183 (phospho-GPR183 Antibody)
Threonine347/Threonine349 (T347/T349) is major phosphorylation site of human GPR183 receptor. The pT347/pT349-GPR183 antibody detects phosphorylation in response to agonists. T347/T349 phosphorylation is likely to be involved in...
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Agonist-induced Serine346/Serine347 phosphorylation of the Angiotensin Receptor 1
pS346/pS347-AT1 (phospho-Angiotensin Receptor 1...
Serine346/Serine347 (S346/S347) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS346/pS347-AT1 antibody detects phosphorylation in response to agonists. S346/S347 phosphorylation is likely to be involved in...
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Agonist-induced Serine335/Threonine336 phosphorylation of the Angiotensin Receptor 1
pS335/pT336-AT1 (phospho-Angiotensin Receptor 1...
Serine335/Threonine336 (S335/T336) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS335/pT336-AT1 antibody detects phosphorylation in response to agonists. S335/T336 phosphorylation is likely to be involved in...
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Agonist-induced Serine331/Threonine332 phosphorylation of the Angiotensin Receptor 1
pS331/pT332-AT1 (phospho-Angiotensin Receptor 1...
Serine331/Threonine332 (S331/T332) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS331/pT332-AT1 antibody detects phosphorylation in response to agonists. S331/T332 phosphorylation is likely to be involved in...
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Validation of the β2-Adrenoceptor in transfected HEK293 cells
β2 (GP-non-phospho), β2-Adrenoceptor Antibody,...
The non-phospho- β2 antibody is directed against the third intracellular loop of human β2 . It can be used to detect total β2 receptors in Western blots independent of phosphorylation. The non-phospho- β2 antibody can also be used to...
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DOP (GP-non-phospho) δ-Opioid Receptor Antibody, Guinea Pig
DOP (GP-non-phospho) δ-Opioid Receptor...
The non-phospho-δ-opioid receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human DOP. It can be used to detect total DOP receptors in Western blots independent of phosphorylation.
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Immunohistochemical identification of Complement C5a Receptor 1 in Spleen
C5a1 (GP-IHC-grade), Complement C5a Receptor 1...
The C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Complement 5a Receptor 1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The C5a1...
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Validation of the Complement C5a Receptor 1 in transfected HEK293 cells
C5a1 (GP-non-phospho), Complement C5a Receptor...
The non-phospho-C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human C5a1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The non-phospho-C5a1...
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KO-Validated
Western blot analysis of G Protein-coupled Receptor 20 in mouse tissues in vivo.
mGPR20 (non-phospho-G Protein-Coupled Receptor...
The GPR20 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse GPR20. It can be used to detect total GPR20 receptors in Western blots independent of phosphorylation. The GPR20 antibody can also be...
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KO-Validated
Immunohistochemical identification of G Protein-coupled Receptor 20 in mouse spleen.
mGPR20 (IHC-grade), G Protein-Coupled Receptor...
The GPR20 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse GPR20. It can be used to detect total GPR20 receptors in Western blots independent of phosphorylation. The GPR20 antibody can also be...
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Agonist-induced Serine401 phosphorylation of the Neurotensin Receptor
pS401-NTS1 (phospho-Neurotensin Receptor 1...
Serine401 (S401) is major phosphorylation site of the Neurotensin Receptor 1 (NTS1). The pS401-NTS1 antibody detects phosphorylation in response to agonists. S401 phosphorylation is likely to be involved in efficient ligand sequestration...
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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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