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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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SAMPLE PACK
Agonist-induced Threonine370 phosphorylation of the µ-opioid receptor
MOP Sample Pack (phospho- and...
MOP Sample Pack consisting of all four available phospho- and one non-phospho-µ-Opioid Receptor Antibodies 5 x 20 µL trial size each. Specifically, this sample pack contains the following antibodies pT370-MOP (7TM0319B), pS375-MOP...
$ 300.00 *
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Agonist-induced Serine320/Serine321 phosphorylation of the CXC Chemokine Receptor 6
pS320/pS321-CXCR6 (phospho-CXC Chemokine...
Serine320/Serine321 (S320/S321) is a major phosphorylation site of the CXCR6 receptor. The pS320/pS321-CXCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S320/S321...
$ 375.00 *
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Agonist-induced Serine325/Threonine327 phosphorylation of the CXC Chemokine Receptor 6
pS325/pT327-CXCR6 (phospho-CXC Chemokine...
Serine325/Threonine327 (S325/T327) is a major phosphorylation site of the CXCR6 receptor. The pS325/pT327-CXCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S325/T327...
$ 375.00 *
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Agonist-induced Serine329/Serine331 phosphorylation of the CXC Chemokine Receptor 6
pS329/pS331-CXCR6 (phospho-CXC Chemokine...
Serine329/Serine331 (S329/S331) is a major phosphorylation site of the CXCR6 receptor. The pS329/pS331-CXCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S329/S331...
$ 375.00 *
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Agonist-induced Threonine367/Serine368 phosphorylation of the CXC Chemokine Receptor 5
pT367/pS368-CXCR5 (phospho-CXC Chemokine...
Threonine367/Serine368 (T367/S368) is a major phosphorylation site of the CXCR5 receptor. The pT367/pS368-CXCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T367/S368...
$ 375.00 *
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Agonist-induced Threonine370/Threonine371 phosphorylation of the CXC Chemokine Receptor 5
pT370/pT371-CXCR5 (phospho-CXC Chemokine...
Threonine370/Threonine371 (T370/T371) is a major phosphorylation site of the CXCR5 receptor. The pT370/pT371-CXCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T370/T371...
$ 375.00 *
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Agonist-induced Threonine347 phosphorylation of the CXC Chemokine Receptor 2
pT347-CXCR2 (phospho-CXC Chemokine Receptor 2...
Threonine347 (T347) is a major phosphorylation site of the CXCR2 receptor. The pT347-CXCR2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T347 phosphorylation is a key...
$ 375.00 *
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Agonist-induced Serine351/Serine352 phosphorylation of the CXC Chemokine Receptor 2
pS351/pS352-CXCR2 (phospho-CXC Chemokine...
Serine351/Serine352 (S351/S352) is a major phosphorylation site of the CXCR2 receptor. The pS351/pS352-CXCR2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S351/S352...
$ 375.00 *
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Agonist-induced Serine353/Threonine356 phosphorylation of the CXC Chemokine Receptor 2
pS353/pT356-CXCR2 (phospho-CXC Chemokine...
Serine353/Threonine356 (S353/T356) is a major phosphorylation site of the CXCR2 receptor. The pS353/pT356-CXCR2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S353/T356...
$ 375.00 *
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Agonist-induced Threonine353 phosphorylation of the Chemokine Receptor 10
pT353-CCR10 (phospho-Chemokine Receptor 10...
Threonine353 (T353) is a major phosphorylation site of the CCR10 receptor. The pT353-CCR10 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T353 phosphorylation is a key regulator...
$ 375.00 *
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Agonist-induced Threonine355 phosphorylation of the Chemokine Receptor 10
pT355-CCR10 (phospho-Chemokine Receptor 10...
Threonine355 (T355) is a major phosphorylation site of the CCR10 receptor. The pT355-CCR10 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T355 phosphorylation is a key regulator...
$ 375.00 *
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Agonist-induced Serine357 phosphorylation of the Chemokine Receptor 6
pS357-CCR6 (phospho-Chemokine Receptor 6 Antibody)
Serine357 (S357) is a major phosphorylation site of the CCR6 receptor. The pS357-CCR6 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S357 phosphorylation is a key regulator of...
$ 375.00 *
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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...
$ 375.00 *
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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...
$ 375.00 *
Details
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Agonist-induced Serine336/Serine337 phosphorylation of the Chemokine Receptor 5
pS336/pS337-CCR5 (phospho-Chemokine Receptor 5...
Serine336/Serine337 (S336/S337) is a major phosphorylation site of the CCR5 receptor. The pS336/pS337-CCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S336/S337...
$ 375.00 *
Details
NEW
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...
$ 375.00 *
Details
8 From 18

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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