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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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Immunohistochemical identification of NPSR in Duodenum
NPSR (IHC-grade), Neuropeptide S Receptor Antibody
The NPSR antibody is directed against the distal end of the carboxyl-terminal tail of human Neuropeptide S Receptor. It can be used to detect total NPS receptors in Western blots independent of phosphorylation. The NPSR antibody can also...
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Agonist-induced Serine339/Serine340 phosphorylation of the Chemokine Receptor CX3CR1
pS339/pS340-CX3CR1 (phospho-CX3CR1 Chemokine...
Serine339/Serine340 (S339/S340) is a major phosphorylation site of the CX3CR1 receptor. The pS339/pS340-CX3CR1 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S339/S340...
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pS327/pS329-CX3CR1 (phospho-CX3CR1 Chemokine Receptor Antibody)
pS327/pS329-CX3CR1 (phospho-CX3CR1 Chemokine...
Serine327/Serine329 (S327/S329) is a major phosphorylation site of the CX3CR1 receptor. The pS327/pS329-CX3CR1 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S327/S329...
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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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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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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...
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Agonist-induced Serine326 phosphorylation of the Cannabinoid Receptor 2
pS326-CB2 (phospho-Cannabinoid Receptor 2...
Serine326 (S326) is major phosphorylation site of the Cannabinoid Receptor 2 (CB2). The pS326-CB2 antibody detects phosphorylation in response to agonists. S326 phosphorylation is likely to be involved in efficient ligand sequestration...
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Agonist-induced Serine351 phosphorylation of the Apelin Receptor
pS351-APLNR (phospho-Apelin Receptor Antibody)
Serine351 (S351) is major phosphorylation site of the Apelin Receptor (APLNR). The pS351-APLNR antibody detects phosphorylation in response to agonists. S351 phosphorylation is likely to be involved in efficient ligand sequestration by...
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Agonist-induced Serine319/Serine322 phosphorylation of the H2 Histamine Receptor
pS319/pS322-H2 (phospho-H2 Histamine Receptor...
Serine319/Ser322 are major phosphorylation site of the H2 histamine receptor. The pS319/pS322-H2 antibody detects phosphorylation in response to high-efficacy agonists. S319/S322 phosphorylation is a key regulator of H2 desensitization,...
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Agonist-induced Serine350 phosphorylation of the Vasopressin Receptor 2
pS350-V2 (phospho-Vasopressin Receptor 2 Antibody)
Serine350 (S350) is major phosphorylation site of the Vasopresssin Receptor 2 (V2). The pS350-V2 antibody detects phosphorylation in response to agonists. S350 phosphorylation is likely to be involved in efficient ligand sequestration by...
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Agonist-induced Threonine359/Threonine360 phosphorylation of the Vasopressin Receptor 2.
pT359/pT360-V2 (phospho-Vasopressin Receptor 2...
Threonine359/Threonine360 (T359/T360) is major phosphorylation site of the Vasopressin Receptor 2 (V2). The pT359/pT360-V2 antibody detects phosphorylation in response to agonists. T359/T360 phosphorylation is likely to be involved in...
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Agonist-induced Serine362/Serine363/Serine364 phosphorylation of the Vasopressin Receptor 2
pS362/pS363/pS364-V2 (phospho-Vasopressin...
Serine362/Serine363/Serine364 (S362/S363/S364) is major phosphorylation site of the Vasopresssin Receptor 2 (V2). The pS362/pS363/pS364-V2 antibody detects phosphorylation in response to agonists. S362/S363/S364 phosphorylation is likely...
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Agonist-induced Threonine378/Serine380 phosphorylation of the Vasopressin Receptor 1A
pT378/pS380-V1A (phospho-Vasopressin Receptor...
Threonine378/Serine380 (T378/S380) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pT378/pS380-V1A antibody detects phosphorylation in response to agonists. T378/S380 phosphorylation is likely to be involved in...
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Agonist-induced Threonine386/Serine389 phosphorylation of the Vasopressin Receptor 1A
pT386/pS389-V1A (phospho-Vasopressin Receptor...
Threonine386/Serine389 (T368/S389) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pT386/pS389-V1A antibody detects phosphorylation in response to agonists. T386/S389 phosphorylation is likely to be involved in...
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Agonist-induced Serine397/Threonine398 phosphorylation of the Vasopressin Receptor 1A
pS397/pT398-V1A (phospho-Vasopressin Receptor...
Serine397/Threonine398 (S397/T398) is major phosphorylation site of the Vasopressin Receptor 1A (V1A). The pS397/pT398-V1A antibody detects phosphorylation in response to agonists. S397/T398 phosphorylation is likely to be involved in...
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pS373/pS374-V1B (phospho-Vasopressin Receptor 1B Antibody)
pS373/pS374-V1B (phospho-Vasopressin Receptor...
Serine373/Serine374 (S373/S374) is major phosphorylation site of the Vasopressin Receptor 1B (V1B). The pS373/pS374-V1B antibody detects phosphorylation in response to agonists. S373/S374 phosphorylation is likely to be involved in...
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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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