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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 Serine355/Serine360 phosphorylation of the Atypical Chemokine Receptor 3/CXC Chemokine Receptor 7
pS355/pS360-ACKR3 (phospho-Atypical Chemokine...
Serine355/Serine360 (S355/S360) is major phosphorylation site of the Atypical Chemokine Receptor 3 (ACKR3, previously called CXCR7). The pS355/pS360-ACKR3 antibody detects phosphorylation in response to agonists. S355/S360...
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Validation of the SUCNR1 Receptor in transfected HEK293 cells
SUCNR1/GPR91 (non-phospho), Succinate Receptor...
The non-phospho-SUCNR1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human SUCNR1. It can be used to detect total SUCNR1 receptors in Western blots independent of phosphorylation. The SUCNR1...
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Validation of the EP2 Receptor in transfected HEK293 cells
EP2 (non-phospho), EP2 Prostanoid Receptor...
The non-phospho-EP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human EP2. It can be used to detect total EP2 receptors in Western blots independent of phosphorylation.The EP2 antibody can also...
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Validation of the EP4 Receptor in transfected HEK293 cells.
EP4 (non-phospho), EP4 Prostanoid Receptor...
The non-phospho-EP4 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human EP4. It can be used to detect total EP4 receptors in Western blots independent of phosphorylation.
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Immunohistochemical identification of Tachykinin Receptor 1 in pancreatic carcinoma
NK1 (IHC-grade), Tachykinin Receptor 1 Receptor...
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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Agonist-induced Threonine339/Threonine341 phosphorylation of NK2 Receptor.
pT339/pT341-NK2 (phospho-Tachykinin Receptor 2...
Threonine339/Threonine341 (T339/T341) is a major phosphorylation site of the NK2 receptor. The pT339/pT341-NK2 antibody detects phosphorylation in response to high-efficacy agonists. T339/T341 phosphorylation is a key regulator of NK2...
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Agonist-induced Serine345/Threonine346 phosphorylation of NK2 Receptor.
pS345/pT346-NK2 (phospho-Tachykinin Receptor 2...
Serine345/Threonine346 (S345/T346) is a major phosphorylation site of the NK2 receptor. The pS345/pT346-NK2 antibody detects phosphorylation in response to high-efficacy agonists. S345/T346 phosphorylation is a key regulator of NK2...
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Agonist-induced Threonine353/Threonine356 phosphorylation of NK2 Receptor.
pT353/pT356-NK2 (phospho-Tachykinin Receptor 2...
Threonine353/Threonine356 (T353/T356) is a major phosphorylation site of the NK2 receptor. The pT353/pT356-NK2 antibody detects phosphorylation in response to high-efficacy agonists. T353/T356 phosphorylation is a key regulator of NK2...
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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 MRGPRX2 Receptor in transfected HEK293 cells
MRGPRX2 (GP-non-phospho), MRGPRX2 Mas-related...
The non-phospho-MRGPRX2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human MRGPRX2. It can be used to detect total MRGPRX2 receptors in Western blots independent of phosphorylation. The MRGPRX2...
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Immunohistochemical identification of MRGPRX2 Receptor in Pancreas
MRGPRX2 (GP-IHC-grade), MRGPRX2 Mas-related...
The non-phospho-MRGPRX2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human MRGPRX2. It can be used to detect total MRGPRX2 receptors in Western blots independent of phosphorylation. The MRGPRX2...
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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 Threonine335 phosphorylation of FPR2 Receptor
pT335-FPR2 (phospho-Formylpeptide Receptor 2...
Threonine335 (T335) is a major phosphorylation site of the FPR2 receptor. The pT335-FPR2 antibody detects phosphorylation in response to high-efficacy agonists. T335 phosphorylation is a key regulator of FPR2 desensitization, β-arrestin...
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Agonist-induced Threonine334/Serine335 phosphorylation of FPR3 Receptor
pT334/pS335-FPR3 (phospho-Formylpeptide...
Threonine334/Serine335 (T334/S335) is a major phosphorylation site of the FPR3 receptor. The pT334/pS335-FPR3 antibody detects phosphorylation in response to high-efficacy agonists. T334/S335 phosphorylation is a key regulator of FPR3...
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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 Serine338/Threonine339 phosphorylation of Formylpeptide Receptor 1
pS338/pT339-FPR1 (phospho-Formylpeptide...
Serine338/Threonine339 (S338/T339) is a major phosphorylation site of the Formylpeptide Receptor 1 (FPR1). The pS338/pT339-FPR1 antibody detects phosphorylation in response to high-efficacy agonists. S338/T339 phosphorylation is a key...
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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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