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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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gonist-induced Threonine357/Serine360 phosphorylation of the Neuropeptide Y receptor 1
pT357/pS360-NPY1 (phospho-Neuropeptide Y...
Threonine357/Serine360 (T357/S360) is a major phosphorylation site of the Neuropeptide Y Receptor 1. The pT357/pS360-NPY1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Validation of the CC Chemokine Receptor 5 in transfected HEK293 cells
CCR5 (GP-non-phospho) Chemokine Receptor 5...
The non-phospho-CCR5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CCR5. It can be used to detect total CCR5 receptors in Western blots independent of phosphorylation. The CCR5 antibody can...
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Agonist-induced Threonine340 phosphorylation of the Chemokine Receptor 5
pT340-CCR5 (phospho-Chemokine Receptor 5 Antibody)
Threonine340 (T340) is a major phosphorylation site of the CCR5 receptor. The pT340-CCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T340 phosphorylation is a key regulator of...
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Agonist-induced Serine429 phosphorylation of the Cannabinoid Receptor 1
pS429-CB1 (phospho-Cannabinoid Receptor 1...
Serine429 (S429) is major phosphorylation site of the Cannabinoid Receptor 1 (CB1). The pS429-CB1 antibody detects phosphorylation in response to agonists. S429 phosphorylation is likely to be involved in efficient ligand sequestration...
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Agonist-induced Serine441 phosphorylation of the Cannabinoid Receptor 1
pS441-CB1 (phospho-Cannabinoid Receptor 1...
Serine441 (S441) is major phosphorylation site of the Cannabinoid Receptor 1 (CB1). The pS441-CB1 antibody detects phosphorylation in response to agonists. S441 phosphorylation is likely to be involved in efficient ligand sequestration...
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Immunohistochemical Identification of Agonist-induced Serine425 phosphorylation of the Cannabinoid Receptor 1 in mouse brain
pS425-CB1 (IHC-grade phospho-Cannabinoid...
Serine425 (S425) is major phosphorylation site of the Cannabinoid Receptor 1 (CB1). The pS425-CB1 antibody detects phosphorylation in response to agonists. S425 phosphorylation is likely to be involved in efficient ligand sequestration...
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Agonist-induced Serine329 phosphorylation of the A2A Adenosine Receptor
pS329-A2A (phospho-A2A Adenosine Receptor...
Serine329 is a major phosphorylation site of the A2A Adenosine receptor. The pS329-A2A antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S329 phosphorylation is a key regulator of A2A...
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pT442/pS443-GIPR (phospho-Gastric Inhibitory Polypeptide Receptor Antibody)
pT442/pS443-GIPR (phospho-Gastric Inhibitory...
Threonine442/Serine443 (T442/S443) is major phosphorylation site of human GIP receptor. The pT442/pS443-GIPR antibody detects phosphorylation in response to agonists. T442/S443 phosphorylation is likely to be involved in efficient ligand...
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Agonist-induced Serine433/Serine435 phosphorylation of GIPR
pS433/pS435-GIPR (phospho-Gastric Inhibitory...
Serine433/Serine435 (S433/S435) is major phosphorylation site of human GIP receptor. The pS433/pS435-GIPR antibody detects phosphorylation in response to agonists. S433/S435 phosphorylation is likely to be involved in efficient ligand...
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Immunohistochemical control of anti-NPS (Neuropeptide S) antibody in mouse coronal sections
NPS (IHC-grade), Neuropeptide S Antibody
The NPS antibody is directed against human Neuropeptide S. The NPS antibody can be used to detect Neuropeptide S in formalin-fixed, paraffin-embedded tissue sections by immunohistochemistry.
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Immunohistochemical identification of Nociceptin/Orphanin FQ in mouse dorsal horn
N/OFQ (IHC-grade), Nociceptin/Orphanin FQ Antibody
The N/OFQ receptor antibody is directed against human Nociceptin/Orphanin FQ. The N/OFQ antibody can be used to detect N/OFQ in formalin-fixed, paraffin-embedded tissue sections by immunohistochemistry.
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Immunohistochemical identification of G Protein-coupled Receptor 84 in lymph node
GPR84 (IHC-grade), G Protein-Coupled Receptor...
The GPR84 antibody is directed against the distal end of the carboxyl-terminal tail of human GRP84. It can be used to detect total GPR84 receptors in Western blots independent of phosphorylation. The GPR84 antibody can also be used to...
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Validation of the δ-Opioid Receptor in transfected HEK293 cells
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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Agonist-induced Serine335/Serine337 phosphorylation of the Sphingosine 1-Phosphate Receptor 3
pS335/pS337-S1P3 (phospho-Sphingosine...
Serine335/Serine337 (S335/S337) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 3 (S1P3). The pS335/pS337-S1P3 antibody detects phosphorylation in response to agonists. S335/S337 phosphorylation is likely to be...
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Immunohistochemical identification of G Protein-coupled Receptor 17 in oligodendrocytes
GPR17 (GP-IHC-grade), G Protein-Coupled...
The GPR17 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP17. It can be used to detect total GPR17 receptors in Western blots independent of phosphorylation. The GPR17 antibody can...
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Immunohistochemical identification of the Glucagon Receptor in pancreas
GCGR (IHC-grade), Glucagon Receptor Antibody
The GCGR antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GCGR. It can be used to detect total GCGR receptors in Western blots independent of phosphorylation. The GCGR antibody can also be...
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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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