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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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Citations
pS337/pT341-SST3 (phospho-Somatostatin Receptor 3 Antibody)
pS337/pT341-SST3 (phospho-Somatostatin Receptor...
Serine337/Threonine341 (S337/T341) is a major phosphorylation site of the somatostatin receptor 3 (SST3). The pS337/pT341-SST3 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Immunohistochemical identification of Somatostatin Receptor 1 in human pituitary.
SST1 (IHC-grade), Somatostatin Receptor 1 Antibody
The SST1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Somatostatin Receptor 1. It can be used to detect total SST1 receptors in Western blots independent of phosphorylation. The SST1...
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KO-Validated
Validation of the Somatostatin Receptor 5 in transfected HEK293 cells.
SST5 (non-phospho), Somatostatin Receptor 5...
The non-phospho-SST5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human SST5. In can be used to detect total SST5 receptors in Western blots independent of phosphorylation. The non-phospho-SST5...
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Figure 1. Agonist-induced Serine375 phosphorylation of the µ-opioid receptor.
pS375-MOP (phospho-µ-Opioid Receptor Antibody)
Serine375 (S375) is the primary phosphorylation site in a hierarchical phosphorylation cascade. The pS375-MOP antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S375...
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Immunohistochemical identification of Bombesin receptor 3 in human ovarian carcinoma.
BB3 (IHC-grade), Bombesin Receptor 3 Antibody
The BB3 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Bombesin Receptor 3. It can be used to detect BB3 in formalin-fixed, paraffin-embedded tissue sections by immunohistochemistry.
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KO-Validated
Validation of the Nociceptin/Orphanin FQ Receptor in transfected HEK293 cells.
NOP (non-phospho), Nociceptin/Orphanin FQ...
The non-phospho-NFQ receptor (NOP) antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human NOP. It can be used to detect total NOP receptors in Western blots independent of phosphorylation. The...
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Validation of the κ-Opioid Receptor in transfected HEK293 cells.
KOP (non-phospho) κ-Opioid Receptor Antibody
The non-phospho-kappa-opioid receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human KOP. It can be used to detect total KOP receptors in Western blots independent of phosphorylation.
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Validation of the Galanin Receptor 1 in transfected HEK293 cells
GAL1 (non-phospho), Galanin Receptor 1 Antibody
The non-phospho-GAL1 receptor antibody is directed against the carboxyl-terminal tail of human GAL1. It can be used to detect total GAL1 receptors in Western blots independent of phosphorylation. The non-phospho-GAL1 antibody can also be...
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NEW
Validation of the Formylpeptide Receptor 3 in transfected HEK293 cells
FPR3 (non-phospho), Formylpeptide Receptor 3...
The FPR3 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Formylpeptide Receptor 3. It can be used to detect total FPR3 receptors in Western blots independent of phosphorylation. The FPR3...
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Agonist-induced Serine422/Serine425 phosphorylation of VPAC1 Receptor
pS422/pS425-VPAC1 (phospho-VIP Receptor 1...
Serine422/Serine425 (S422/S425) is a major phosphorylation site of the VPAC1 receptor. The pS422/pS425-VPAC1 antibody detects phosphorylation in response to high-efficacy agonists. S422/S425 phosphorylation is a key regulator of VPAC1...
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SAMPLE PACK
Agonist-induced Serine324/Serine325 phosphorylation of the CXC Chemokine Receptor 4.
CXCR4 Sample Pack (phospho- and...
CXCR4 Sample Pack consisting of all five available phospho- and non-phospho-CXCR4 Antibodies 5 x 20 µL trial size each. Specifically, this sample pack contains the following antibodies pS324/pS325-CXCR4 (7TM0071A), pS330-CXCR4...
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pS417/pS419-CCK1 (phospho-Cholecystokinin Receptor 1 Antibody)
pS417/pS419-CCK1 (phospho-Cholecystokinin...
Serine417/Serine419 (S417/S419) is a major phosphorylation site of the CCK1 receptor. The pS417/pS419-CCK1 antibody detects phosphorylation in response to high-efficacy agonists. S417/S419 phosphorylation is a key regulator of CCK1...
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Citations
Agonist-induced Threonine362/Serine363 phosphorylation of the Nociceptin/Orphanin FQ Receptor.
pT362/pS363-NOP (phospho-Nociceptin/Orphanin FQ...
Threonine362/Serine363 (T362/S363) is a major phosphorylation site of the nociceptin/orphanin FQ peptide receptor (NFQ receptor, NOP). The pT362/pS363-NOP antibody detects phosphorylation in response to agonists but not after PKC...
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Agonist-induced Threonine356/Threonine357 phosphorylation of NK1 Receptor
pT356/pT357-NK1 (phospho-Tachykinin Receptor 1...
Threonine356/Threonine357 (T356/T357) is a major phosphorylation site of the NK1 receptor. The pT356/pT357-NK1 antibody detects phosphorylation in response to high-efficacy agonists. T356/T357 phosphorylation is a key regulator of NK1...
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Agonist-induced Serine338/Threonine339 phosphorylation of NK1 Receptor
pS338/pT339-NK1 (phospho-Tachykinin Receptor 1...
Serine338/Threonine339 (S338/T339) is a major phosphorylation site of the NK1 receptor. The pS338/pT339-NK1 antibody detects phosphorylation in response to high-efficacy agonists. S338/T339 phosphorylation is a key regulator of NK1...
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pT344/pS347-NK1 (phospho-Tachykinin Receptor 1 Antibody)
pT344/pS347-NK1 (phospho-Tachykinin Receptor 1...
Threonine344/Serine347 (T344/S347) is a major phosphorylation site of the NK1 receptor. The pT344/pS347-NK1 antibody detects phosphorylation in response to high-efficacy agonists. T344/S347 phosphorylation is a key regulator of NK1...
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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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