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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 G protein-coupled receptor 87 in transfected HEK293 cells.
GPR87 (non-phospho-G Protein-Coupled Receptor...
The GPR87 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP87. It can be used to detect total GPR87 receptors in Western blots independent of phosphorylation. The GPR87 antibody can...
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Agonist-induced Threonine404 phosphorylation of the β1-Adrenoceptor
pT404-β1 (phospho-β1-Adrenoceptor Antibody)
Threonine404 (T404) is a major phosphorylation site of the β1 adrenoceptor. The pT404-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T404 phosphorylation is primarily...
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Validation of the β1-Adrenoceptor in transfected HEK293 cells
β1 (non-phospho), β1-Adrenoceptor Antibody
The non-phospho- β1 antibody is directed against the carboxyl-terminal tail of human β1 . It can be used to detect total β1 receptors in Western blots independent of phosphorylation. The non-phospho- β1 antibody can also be used to...
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Agonist-induced Serine366/Serine369 phosphorylation of the Proteinase-Activated Receptor 4
pS366/pS369-PAR4 (phospho-Proteinase-Activated...
Serine366/Serine369 (S366/S369) is a major phosphorylation site of the Proteinase-Activated Receptor 4 (PAR4). The pS366/pS369-PAR4 antibody detects phosphorylation in response to agonists. S366/S369 phosphorylation is likely to be...
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Agonist-induced Serine374/Threonine379 phosphorylation of the Proteinase-Activated Receptor 4
pS374/pT379-PAR4 (phospho-Proteinase-Activated...
Serine374/Threonine379 (S374/T379) is a major phosphorylation site of the Proteinase-Activated Receptor 4 (PAR4). The pS374/pT379-PAR4 antibody detects phosphorylation in response to agonists. S374/T379 phosphorylation is likely to be...
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Agonist-induced Serine381/Serine382 phosphorylation of the Proteinase-Activated Receptor 4
pS381/pS382-PAR4 (phospho-Proteinase-Activated...
Serine381/Serine382 (S381/S382) is a major phosphorylation site of the Proteinase-Activated Receptor 4 (PAR4). The pS381/pS382-PAR4 antibody detects phosphorylation in response to agonists. S381/S382 phosphorylation is likely to be...
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Immunohistochemical Identification of Serine296/Serine297 phosphorylation of the FFA Receptor 2 in mouse white adipose tissue and peyer's patches.
pS296/pS297-FFA2 (IHC-grade phospho-FFA2 Antibody)
Serine296/Serine297 (S296/S297) is major phosphorylation site of the FFA Receptor 2 (FFA2). The pS296/pS297-FFA2 antibody detects constitutive phosphorylation of FFA2.
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Immunohistochemical identification of GPR15 Receptor in kidney
GPR15 (IHC-grade), G Protein-Coupled Receptor...
The GPR15 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP15. It can be used to detect total GPR15 receptors in Western blots independent of phosphorylation. The GPR15 antibody can...
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Immunohistochemical identification of GPR39 Receptor in Small Intestine
GPR39 (IHC-grade), G Protein-Coupled Receptor...
The GPR39 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP39. It can be used to detect total GPR39 receptors in Western blots independent of phosphorylation. The GPR39 antibody can...
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Immunohistochemical identification of NPSR in Duodenum
NPSR (IHC-grade), Neuropeptide S Receptor Antibody
The NPSR antibody is directed against the distal end of the carboxyl-terminal tail of human Neuropeptide S Receptor. It can be used to detect total NPS receptors in Western blots independent of phosphorylation. The NPSR antibody can also...
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Agonist-induced Serine339/Serine340 phosphorylation of the Chemokine Receptor CX3CR1
pS339/pS340-CX3CR1 (phospho-CX3CR1 Chemokine...
Serine339/Serine340 (S339/S340) is a major phosphorylation site of the CX3CR1 receptor. The pS339/pS340-CX3CR1 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S339/S340...
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Agonist-induced Serine351 phosphorylation of the Apelin Receptor
pS351-APLNR (phospho-Apelin Receptor Antibody)
Serine351 (S351) is major phosphorylation site of the Apelin Receptor (APLNR). The pS351-APLNR antibody detects phosphorylation in response to agonists. S351 phosphorylation is likely to be involved in efficient ligand sequestration by...
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Agonist-induced Serine319/Serine322 phosphorylation of the H2 Histamine Receptor
pS319/pS322-H2 (phospho-H2 Histamine Receptor...
Serine319/Ser322 are major phosphorylation site of the H2 histamine receptor. The pS319/pS322-H2 antibody detects phosphorylation in response to high-efficacy agonists. S319/S322 phosphorylation is a key regulator of H2 desensitization,...
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Agonist-induced Threonine271 phosphorylation of the M2 Muscarinic Acetycholine Receptor
pT271-M2 (phospho-M2 Muscarinic Acetylcholine...
Threonine271 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pT271-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T271 phosphorylation is a key...
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Agonist-induced Serine282/Serine283 phosphorylation of the M2 Muscarinic Acetylcholine Receptor
pS282/pS283-M2 (phospho-M2 Muscarinic...
Serine282/Serine283 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pS282/pS283-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S282/S283...
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Agonist-induced Threonine307/Serine309 phosphorylation of the M2 Muscarinic Acetylcholine Receptor
pT307/pS309-M2 (phospho-M2 Muscarinic...
Threonine307/Serine309 is a major phosphorylation site of the M2 Muscarinic Acetylcholine Receptor. The pT307/pS309-M2 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T307/S309...
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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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