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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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pT360/pS361-CCR6 (phospho-Chemokine Receptor 6 Antibody)
pT360/pS361-CCR6 (phospho-Chemokine Receptor 6...
Threonine360/Serine361 (T360/S361) is a major phosphorylation site of the CCR6 receptor. The pT360/pS361-CCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T360/S361...
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pT363-CCR6 (phospho-Chemokine Receptor 6 Antibody)
pT363-CCR6 (phospho-Chemokine Receptor 6 Antibody)
Threonine363 (T363) is a major phosphorylation site of the CCR6 receptor. The pT363-CCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T363 phosphorylation is a key regulator of...
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Immunohistochemical identification of Complement C5a Receptor 1 in human spleen.
C5a1 (IHC-grade), Complement C5a Receptor 1...
The C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Complement 5a Receptor 1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The C5a1...
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Immunohistochemical identification of Complement C5a Receptor 1 in Spleen
C5a1 (GP-IHC-grade), Complement C5a Receptor 1...
The C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Complement 5a Receptor 1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The C5a1...
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pS296/pS297-M4 (phospho-M4 Muscarinic Acetylcholine Receptor Antibody)
pS296/pS297-M4 (phospho-M4 Muscarinic...
Serine296/Serine297 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS296/pS297-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S296/S297...
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pT311/pS314-M4 (phospho-M4 Muscarinic Acetylcholine Receptor Antibody)
pT311/pS314-M4 (phospho-M4 Muscarinic...
Threonine311/Serine314 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pT311/pS314-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T311/S314...
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Validation of the Smoothened Receptor in transfected HEK293 cells
SMO (non-phospho) Smoothened Receptor Antibody
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Validation of the Smoothened Receptor in transfected HEK293 cells
SMO (GP-non-phospho) Smoothened Receptor...
The non-phospho SMO receptor antibody is directed against the distal end of the carboxyl-terminal tail human SMO receptor. It can be used to detect total SMO receptors in Western blots independent of phosphorylation. It can also be used...
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Validation of the Melatonin Receptor 1 in transfected HEK293 cells
MT1 (non-phospho) Melatonin Receptor 1 Antibody
The non-phospho MT1 receptor antibody is directed against the distal end of the carboxyl-terminal tail human MT1 receptor. It can be used to detect total MT1 receptors in Western blots independent of phosphorylation. It can also be used...
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Agonist-induced Serine355/Serine356 phosphorylation of the β2 Adrenoceptor
pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)
Serine355/Serine356 (S355/S356) is a major phosphorylation site of the β2 adrenoceptor. The pS355/pS356-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S355/S356...
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Immunohistochemical Identification of Serine594/Threonine597/Serine599 phosphorylation of the SMO Receptor in primary cilia in the mouse neural tube
pS594/pT597/pS599-SMO (phospho-SMO Receptor...
Serine594/Threonine597/Serine599 (S594/T597/S599) is major GRK2 phosphorylation site of the SMO receptor. The pS594/pT597/pS599-SMO antibody detects phosphorylation in response to agonists. The pS594/pT597/pS599-SMO antibody can be used...
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Agonist-induced Serine594/Threonine597/Serine599 phosphorylation of the SMO Receptor
pS594/pT597/pS599-SMO (phospho-SMO Receptor...
Serine594/Threonine597/Serine599 (S594/S597/S599) is major GRK2 phosphorylation site of the SMO receptor. The pS594/pT597/pS599-SMO antibody detects phosphorylation in response to agonists. The pS594/pT597/pS599-SMO antibody can be used...
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Agonist-induced Serine351/Serine354 phosphorylation of the Neuropeptide Y
pS351/pS354-NPY2 (phospho-Neuropeptide Y...
Serine351/Serine354 (S351/S354) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS351/pS354-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Agonist-induced Serine354/Threonine356 phosphorylation of the Neuropeptide Y receptor 2
pS354/pT356-NPY2 (phospho-Neuropeptide Y...
Serine354Threonine356 (S354/T356) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS354/pT356-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Agonist-induced Serine369/Serine374 phosphorylation of the Neuropeptide Y receptor 2
pS369/pS374-NPY2 (phospho-Neuropeptide Y...
Serine369/Serine374 (S369/S374) is a major phosphorylation site of the Neuropeptide Y Receptor 2. The pS369/pS374-NPY2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Agonist-induced Threonine362/Serine363 phosphorylation of the Neuropeptide Y receptor 1
pT362/pS363-NPY1 (phospho-Neuropeptide Y...
Threonine362/Serine363 (T362/S363) is a major phosphorylation site of the Neuropeptide Y Receptor 1. The pT362/pS363-NPY1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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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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