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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 the Hydroxycarboxylic Acid Receptor 2 in spleen
HCA2 (GP-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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Validation of the Dopamine Receptor 5 in transfected HEK293 cells
D5 (non-phospho), Dopamine Receptor 5 Antibody
The non-phospho-Dopamine Receptor 5 Antibody is directed against the distal end of the carboxyl-terminal tail of human D5. It can be used to detect total D5 receptors in Western blots independent of phosphorylation. The non-phospho-D5...
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pT354-D1 (phospho-Dopamine Receptor 1 Antibody)
pT354-D1 (phospho-Dopamine Receptor 1 Antibody)
Threonine354 (T354) is a major phosphorylation site of the D1 receptor. The pT354-D1 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T354 phosphorylation is a key regulator of D1...
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Agonist-induced Threonine382/Threonine386 phosphorylation of the Dopamine Receptor 5
pT382/pT386-D5 (phospho-Dopamine Receptor 5...
Threonine382/Threonine386 (T382/T386) is a major phosphorylation site of the D5 receptor. The pT382/pT386-D5 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T382/T386...
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Agonist-induced Serine345 phosphorylation of the Sphingosine 1-Phosphate Receptor 5
pS345-S1P5 (phospho-Sphingosine 1-Phosphate...
Serine345 (S345) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 5 (S1P5). The pS345-S1P5 antibody detects phosphorylation in response to agonists. S345 phosphorylation is likely to be involved in efficient ligand...
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MCH2 (non-phospho) Melanin-Concentrating Hormone Receptor 2 Antibody
MCH2 (non-phospho) Melanin-Concentrating...
The non-phospho MCH2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human MCH2 receptor. It can be used to detect total MCH2 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the Neuropeptide Y Receptor 1 in transfected HEK293 cells
NPY1 (non-phospho) Neuropeptide Y Receptor 1...
The non-phospho NPY1 receptor antibody is directed against the distal end of the carboxyl-terminal tail human NPY1 receptor. It can be used to detect total NPY1 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the Neuropeptide Y Receptor 2 in transfected HEK293 cells
NPY2 (non-phospho) Neuropeptide Y Receptor 2...
The non-phospho NPY2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human NPY2 receptor. It can be used to detect total NPY2 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the QRFP Receptor in transfected HEK293 cells
QRFPR (non-phospho) QRFP Receptor Antibody
The non-phospho QRFP receptor antibody is directed against the distal end of the carboxyl-terminal tail human QRFP receptor. It can be used to detect total QRFP receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the Secretin Receptor in transfected HEK293 cells
SCTR (non-phospho) Secretin Receptor Antibody
The non-phospho SCTR receptor antibody is directed against the distal end of the carboxyl-terminal tail human SCTR receptor. It can be used to detect total SCTR receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the P2RY Purinoceptor 8 in transfected HEK293 cells
P2RY8 (non-phospho), P2RY Purinoceptor 8 Antibody
The non-phospho-P2RY8 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human P2RY8. It can be used to detect total P2RY8 receptors in Western blots independent of phosphorylation. The...
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Agonist-induced Serine344/Serine345 phosphorylation of the P2Y Receptor
pS344/pS345-P2Y4 (phospho-P2Y Purinoceptor 4...
Serine344/Serine345 (S344/S345) is a major phosphorylation site of the P2Y4 receptor. The pS344/pS345-P2Y4 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S344/S345...
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Agonist-induced Serine323 phosphorylation of the P2Y Receptor 12.
pS323-P2Y12 (phospho-P2Y Purinoceptor 12 Antibody)
Serine323 (S323) is a major phosphorylation site of the P2Y12 receptor. The pS323-P2Y12 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S323 phosphorylation is a key regulator of...
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Validation of the P2Y Purinoceptor 12 in transfected HEK293 cells
P2Y12 (non-phospho) P2Y Purinoceptor 12 Antibody)
The non-phospho P2Y12 antibody is directed against the distal end of the carboxyl-terminal tail human P2Y12. It can be used to detect total P2Y12 receptors in Western blots independent of phosphorylation. It can also be used to isolate...
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Validation of the Oxytocin Receptor in transfected HEK293 cells
OT (non-phospho), Oxytocin Receptor Antibody
The non-phospho-OT Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human OT. It can be used to detect total OT receptors in Western blots independent of phosphorylation. The non-phospho-OT antibody...
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Validation of the Formylpeptide Receptor 1 in transfected HEK293 cells
FPR1 (non-phospho), Formylpeptide Receptor 1...
The non-phospho-FPR1 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human FPR1. It can be used to detect total FPR1 receptors in Western blots independent of phosphorylation. The non-phospho-FPR1...
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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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