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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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Validation of the Glucagon-Like Peptide 2 Receptor in transfected HEK293 cells
GLP2 (non-phospho) Glucagon-Like Peptide 2...
The non-phospho GLP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human GLP2 receptor. It can be used to detect total GLP2 receptors in Western blots independent of phosphorylation. It can also be...
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Immunohistochemical identification of Glucagon-Like Peptide 2 Receptor in enteric ganglion
GLP2 (IHC-grade), Glucagon-Like Peptide 2...
The non-phospho GLP2 receptor antibody is directed against the distal end of the carboxyl-terminal tail human GLP2 receptor. It can be used to detect total GLP2 receptors in Western blots independent of phosphorylation. It can also be...
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Validation of the Glucagon Receptor in transfected HEK293 cells
GCGR (non-phospho) Glucagon Receptor Antibody
The non-phospho GCGR antibody is directed against the distal end of the carboxyl-terminal tail of human GCGR. It can be used to detect total GCGR receptors in Western blots independent of phosphorylation. It can also be used to isolate...
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Validation of the Gastric Inhibitory Polypeptide Receptor in transfected HEK293 cells
GIPR (non-phospho) Gastric Inhibitory...
The non-phospho GIPR antibody is directed against the distal end of the carboxyl-terminal tail human GIPR. It can be used to detect total GIPR receptors in Western blots independent of phosphorylation. It can also be used to isolate and...
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Agonist-induced Threonine323/Serine324/Serine325 phosphorylation of the Lysophosphatidic Acid Receptor 2
pT323/pS324/pS325-LPA2...
Threonine323/Serine324/Serine325 (T323/S324/325) is major phosphorylation site of the Lysophosphatidic Acid Receptor 2 (LPA2). pT323/pS324/pS325-LPA2 antibody detects phosphorylation in response to agonists. T323/S324/325 phosphorylation...
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pS331/pT332-LPA2 (phospho-Lysophosphatidic Acid Receptor 2 Antibody)
pS331/pT332-LPA2 (phospho-Lysophosphatidic Acid...
Serine331/Threonine332 (S331/T332) is major phosphorylation site of the Lysophosphatidic Acid Receptor 2 (LPA2). pS331/pT332-LPA2 antibody detects phosphorylation in response to agonists. S331/T332 phosphorylation is likely to be...
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CALCR [UMB26] (RM-IHC-Grade-Calcitonin Receptor-Like Receptor Antibody), Rabbit Monclonal
CALCR [UMB26] (RM-IHC-Grade-Calcitonin...
The non-phospho-CALCR receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CALCR. It can be used to detect total CALCR receptors in Western blots independent of phosphorylation. The CALCR antibody...
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Agonist-induced Serine422/Serine425 phosphorylation of the Calcitonin Recepto
pS422/pS425-CT (phospho-Calcitonin Receptor...
Serine422/Serine425 (S422/S425) is a major phosphorylation site of the Calcitonin receptor. The pS422/pS425-CT antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S422/S425...
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Agonist-induced Serine349/Threonine350/Threonine351 phosphorylation of the Sphingosine 1-Phosphate Receptor 4
pS349/pT350/pT351-S1P4 (phospho-Sphingosine...
Serine349/Threonin350/Threonine351 (S349/T350/T351) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 4 (S1P4). The pS349/pT350/pT351-S1P4 antibody detects phosphorylation in response to agonists. S349/T350/T351...
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Agonist-induced Serine396/Threonine phosphorylation of the Neuromedin U Receptor 1
pS396/pT397-NMU1 (phospho-Neuromedin U Receptor...
Serine396/Threonine397 is major phosphorylation site of the Neuromedin U Receptor 1 (NMU1). The pS396/pT397-NMU1 antibody detects phosphorylation in response to agonists. S396/pT397 phosphorylation is likely to be involved in efficient...
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Agonist-induced Serine445 phosphorylation of Glucagon Receptor
pS445-GCGR (phospho-Glucagon Receptor Antibody)
Serine445 (S445) is major phosphorylation site of human GCGR receptor. The pS445-GCGR antibody detects phosphorylation in response to agonists. S445 phosphorylation is likely to be involved in efficient ligand sequestration by the...
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Immunohistochemical identification of G Protein-coupled Receptor 17 in oligodendrocytes
GPR17 (GP-IHC-grade), G Protein-Coupled...
The GPR17 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP17. It can be used to detect total GPR17 receptors in Western blots independent of phosphorylation. The GPR17 antibody can...
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Validation of the mouse Complement C5a Receptor 1 in transfected HEK293 cells
mC5a1 (GP-non-phospho), Mouse Complement C5a...
The non-phospho-C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse C5a1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The non-phospho-C5a1...
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Validation of the mouse Complement C5a Receptor 2 in transfected HEK293 cells
mC5a2 (GP-non-phospho), Mouse Complement C5a...
The non-phospho-C5a2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of mouse C5a2. It can be used to detect total C5a2 receptors in Western blots independent of phosphorylation. The non-phospho-C5a2...
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Agonist-induced Serine326/Threonine327/Serine328 phosphorylation of the Hydroxycarboxylic Acid Receptor 2
pS326/pT327/pS328-HCA2...
Serine326/Threonine327/Serine328 (S326/T327/S328) is major phosphorylation site of the Hydroxycarboxylic Acid Receptor 2 (HCA2). The pS326/pT327/pS328-HCA2 antibody detects phosphorylation in response to agonists. S326/T327/S328...
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Agonist-induced Serine338/Threonine342/Threonine345 phosphorylation of the M5 Muscarinic Acetylcholine Receptor
pS338/pT342/pT345-M5 (phospho-M5 Muscarinic...
Serine338/Threonine342/Threonine345 is a major phosphorylation site of the M5 Muscarinic Acetylcholine Receptor. The pS338/pT342/pT345-M5 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC...
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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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