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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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pS364/pS366/pS368-S1P4 (phospho-Sphingosine 1-Phosphate Receptor 4 Antibody)
pS364/pS366/pS368-S1P4 (phospho-Sphingosine...
Serine364/Serine366/Serine368 (S364/S366/S368) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 4 (S1P4). The pS364/pS366/pS368-S1P4 antibody detects phosphorylation in response to agonists. S364/S366/S368...
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Agonist-induced Threonine315/Threonine316 phosphorylation of the M4 Muscarinic Acetylcholine Receptor
pT315/pT316-M4 (phospho-M4 Muscarinic...
Threonine315/Threonine316 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pT315/pT316-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T315/T316...
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Validation of the DP1 Prostanoid Receptor in transfected HEK293 cells
DP1 (non-phospho), DP1 Prostanoid Receptor...
The non-phospho-DP1 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human DP1. It can be used to detect total DP1 receptors in Western blots independent of phosphorylation. The non-phospho-DP1...
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Validation of the DP2 Prostanoid Receptor in transfected HEK293 cells
DP2 (non-phospho), DP2 Prostanoid Receptor...
The non-phospho-DP2 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human DP2. It can be used to detect total DP2 receptors in Western blots independent of phosphorylation. The non-phospho-DP2...
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Validation of the Vasopressin Receptor 2 in transfected HEK293 cells
V2 (non-phospho), Vasopressin Receptor 2 Antibody
The non-phospho-V2 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V2. It can be used to detect total V2 receptors in Western blots independent of phosphorylation. The non-phospho-V2 antibody...
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Validation of the Vasopressin Receptor 1B in transfected HEK293 cells.
V1B (non-phospho), Vasopressin Receptor 1B...
The non-phospho-V1B Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1B. It can be used to detect total V1B receptors in Western blots independent of phosphorylation. The non-phospho-V1B...
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Immunohistochemical identification of Serine298/Serine300 phosphorylation of M4 Receptor in striatum
pS298/pS300-M4 (IHC-grade phospho-M4 Muscarinic...
Serine298/Serine300 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pS298/pS300-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S298/S300...
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Validation of the P2Y Purinoceptor 4 in transfected HEK293 cells
P2Y4 (non-phospho), P2Y Purinoceptor 4 Antibody
The non-phospho-P2Y4 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human P2Y4. It can be used to detect total P2Y4 receptors in Western blots independent of phosphorylation. The non-phospho-P2Y4...
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Validation of the Vasopressin Receptor 1A in transfected HEK293 cells
V1A (non-phospho), Vasopressin Receptor 1A...
The non-phospho-V1A Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1A. It can be used to detect total V1A receptors in Western blots independent of phosphorylation. The non-phospho-V1A...
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Agonist-induced Serine350/Serine352 phosphorylation of the 5-Hydroxytryptamine Receptor 6
pS350/pS352-5-HT6 (phospho-5-HT Receptor 6...
Serine350/Serine352 is a major phosphorylation site of the 5-HT Receptor 6 (5-HT6). The pS350/pS352-5-HT6 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S350/S352 phosphorylation is a...
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pT343/pS347/pT348-5-HT4 (phospho-5-HT Receptor 4 Antibody)
pT343/pS347/pT348-5-HT4 (phospho-5-HT Receptor...
Threonine343/Serine347/Threonine348 is a major phosphorylation site of the 5-HT4 receptor. The pT343/pS347/pT348-5-HT4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T343/S347/T348...
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Agonist-induced Serine354/Threonine355 phosphorylation of the 5-Hydroxytryptamine Receptor 4
pS354/pT355-5-HT4 (phospho-5-HT Receptor 4...
Serine354/Threonine355 is a major phosphorylation site of the 5-HT4 receptor. The pS354/pT355-5-HT4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. S354/T355 phosphorylation is a key...
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Validation of the CC Chemokine Receptor 6 in transfected HEK293 cells
CCR6 (non-phospho) Chemokine Receptor 6 Antibody
The non-phospho-CCR6 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CCR6. It can be used to detect total CCR6 receptors in Western blots independent of phosphorylation. The CCR6 antibody can...
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Validation of the CC Chemokine Receptor 10 in transfected HEK293 cells
CCR10 (non-phospho) Chemokine Receptor 10 Antibody
The non-phospho-CCR10 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CCR10. It can be used to detect total CCR10 receptors in Western blots independent of phosphorylation. The CCR10 antibody...
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Agonist-induced Threonine338 phosphorylation of the Hydroxycarboxylic Acid Receptor 2
pT338-HCA2 (phospho-Hydroxycarboxylic Acid...
Threonine338 (T338) is major phosphorylation site of the Hydroxycarboxylic Acid Receptor 2 (HCA2). The pT338-HCA2 antibody detects phosphorylation in response to agonists. T338 phosphorylation is likely to be involved in efficient ligand...
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Immunohistochemical identification of the Hydroxycarboxylic Acid Receptor 2 in liver
HCA2 (IHC-grade), Hydroxycarboxylic Acid...
The non-phospho HCA2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human HCA2 receptor. It can be used to detect total HCA2 receptors in Western blots independent of phosphorylation. It 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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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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