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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 Serine352/Threonine354 phosphorylation of Chemerin Receptor 1
pS352/pT354-CMKLR1 (phospho-Chemerin Receptor 1...
Serine352/Threonine354 (S352/T354) is a major phosphorylation site of the Chemerin receptor 1 (CMKLR1). The pS352/pT354-CMKLR1 antibody detects phosphorylation in response to high-efficacy agonists. S352/T354 phosphorylation is a key...
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Agonist-induced Serine345/Serine346 phosphorylation of Chemerin Receptor 1
pS345/pS346-CMKLR1 (phospho-Chemerin Receptor 1...
Serine345/Serine346 (S345/S346) is a major phosphorylation site of the Chemerin receptor 1 (CMKLR1). The pS345/pS346-CMKLR1 antibody detects phosphorylation in response to high-efficacy agonists. S345/S346 phosphorylation is a key...
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Agonist-induced Serine338/Serine339 phosphorylation of the CXC Chemokine Receptor 4
pS338/pS339-CXCR4 (phospho-CXC Chemokine...
Serine338 and Serine339 (S338/S339) are a major phosphorylation sites of the CXCR4 receptor. The pS338/pS339-CXCR4 antibody detects phosphorylation in response to agonists and after PKC activation. S338/S339 phosphorylation is a key...
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Validation of the M3 Muscarinic Acetylcholine Receptor in transfected HEK293 cells
M3 (non-phospho-M3 Muscarinic Acetylcholine...
The non-phospho-M3 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human M3 Muscarinic Acetylcholine Receptor (M3). It can be used to detect total M3 receptors in Western blots independent of...
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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 Bradykinin Receptor 2 in transfected HEK293 cells
B2 (non-phospho-Bradykinin Receptor 2 Antibody)
The non-phospho-B2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human B2. It can be used to detect total B2 receptors in Western blots independent of phosphorylation. The non-phospho-B2 antibody...
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Agonist-induced Serine330 phosphorylation of the CXC Chemokine Receptor 4
pS330-CXCR4 (phospho-CXC Chemokine Receptor 4...
Serine330(S330) is a major phosphorylation site of the CXCR4 receptor. The pS330-CXCR4 antibody detects phosphorylation in response to agonists and after PKC activation. S330 phosphorylation is a key regulator of CXCR4 desensitization,...
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Agonist-induced Serine362/Serine363 phosphorylation of Ghrelin Receptor
pS362/pS363-GHSR (phospho-Ghrelin Receptor...
Serine362/Serine363 (S362/S363) is major phosphorylation site of the Ghrelin Receptor (GHSR). The pS362/pS363-GHSR antibody detects phosphorylation in response to agonists. S362/S363 phosphorylation is likely to be involved in efficient...
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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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Validation of the PACAP Receptor 1 in transfected HEK293 cells
PAC1 (non-phospho), PACAP Receptor 1 Antibody
The PAC1 antibody is directed against the distal end of the carboxyl-terminal tail of human PACAP Receptor 1. It can be used to detect total PAC1 receptors in Western blots independent of phosphorylation. The PAC1 antibody can also be...
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Validation of the Urotensin Receptor in transfected HEK293 cells
UT (non-phospho), Urotensin Receptor Antibody
The UT receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Urotensin Receptor. It can be used to detect total UT receptors in Western blots independent of phosphorylation. The UT antibody can also...
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Validation of Atypical Chemokine Receptor 4 in transfected HEK293 cells
ACKR4 (non-phospho), Atypical Chemokine...
The ACKR4 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Atypical Chemokine Receptor 4. It can be used to detect total ACKR4 receptors in Western blots independent of phosphorylation. The...
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Validation of the Tachykinin Receptor 1 in transfected HEK293 cells
NK1 (non-phospho), Tachykinin Receptor 1 Antibody
The NK1 antibody is directed against the distal end of the carboxyl-terminal tail of human Tachykinin Receptor 1 (also called Substance P Receptor). It detects selectively the canonical full-length NK1 Tachykinin Receptor. It can be used...
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Validation of the Cholecystokinin Receptor 2 in transfected HEK293 cells
CCK2 (non-phospho), Cholecystokinin Receptor 2...
The CCK2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Cholecystokinin Receptor 2 (previously called CCKB). It can be used to detect total CCK2 receptors in Western blots independent of...
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pT408/pT409-CCK1 (phospho-Cholecystokinin Receptor 1 Antibody)
pT408/pT409-CCK1 (phospho-Cholecystokinin...
Threonine408/Threonine409 (T408/T409) is a major phosphorylation site of the CCK1 receptor. The pT408/pT409-CCK1 antibody detects phosphorylation in response to high-efficacy agonists. T408/T409 phosphorylation is a key regulator of CCK1...
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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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Premium Phosphosite-Specific 7TM Antibodies

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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