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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 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 human GCGR. It can be used to detect total GCGR receptors in Western blots independent of phosphorylation. It can also be used to isolate and...
$ 375.00 *
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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...
$ 375.00 *
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Agonist-induced Serine354/Threonine357/Threonine358 phosphorylation of the Lysophosphatidic Acid Receptor 4
pS354/pT357/pT358-LPA4...
Serine354/Threonine357/Threonine358 (S354/T357/T358) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pS354/pT357/pT358-LPA4 antibody detects phosphorylation in response to agonists. S354/T357/T358...
$ 375.00 *
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Agonist-induced Threonine341/Threonine342/Serine345 phosphorylation of the Lysophosphatidic Acid Receptor 4
pT341/pT342/pS345-LPA4...
Threonine341/Threonine342/Serine345 (T341/T342/S345) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pT341/pT342/pS345-LPA4 antibody detects phosphorylation in response to agonists. T341/T342/S345...
$ 375.00 *
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Agonist-induced Threonine336/Threonine338 phosphorylation of the Lysophosphatidic Acid Receptor 4
pT336/pT338-LPA4 (phospho-Lysophosphatidic Acid...
Threonine336/Threonine338 (T336/T338) is major phosphorylation site of the Lysophosphatidic Acid Receptor 4 (LPA4). The pT336/pT338-LPA4 antibody detects phosphorylation in response to agonists. T336/T338 phosphorylation is likely to be...
$ 375.00 *
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Agonist-induced Threonine338/Serine341 phosphorylation of the Lysophosphatidic Acid Receptor 1
pT338/pS341-LPA1 (phospho-Lysophosphatidic Acid...
Threonine338/Serine341 (T338/S341) is major phosphorylation site of the Lysophosphatidic Acid Receptor 1 (LPA1). The pT338/pS341-LPA1 antibody detects phosphorylation in response to agonists. T338/S341 phosphorylation is likely to be...
$ 375.00 *
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Agonist-induced Serine354/Serine359 phosphorylation of the Relaxin Receptor 4
pS354/pS359-RXFP4 (phospho-Relaxin Receptor 4...
Serine354/Serine359 (S354/S359) is major phosphorylation site of the Relaxin Receptor 4 (RXFP4). The pS354/pS359-RXFP4 antibody detects phosphorylation in response to agonists. S354/S359 phosphorylation is likely to be involved in...
$ 375.00 *
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Agonist-induced Threonine419/Threonine421/Threonine422 phosphorylation of the Relaxin Receptor 3
pT419/pT421/pT422-RXFP3 (phospho-Relaxin...
Threonine419/Threonine421/Threonine422 (T419/T421/T422) is major phosphorylation site of the Relaxin Receptor 3 (RXFP3). The pT419/pT421/pT422-RXFP3 antibody detects phosphorylation in response to agonists. T419/T421/T422 phosphorylation...
$ 375.00 *
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GPR65 (IHC-grade), G Protein-Coupled Receptor 65 Antibody
GPR65 (IHC-grade), G Protein-Coupled Receptor...
The GPR65 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP65. It can be used to detect total GPR65 receptors in Western blots independent of phosphorylation. The GPR65 antibody can...
$ 375.00 *
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GPR61 (IHC-grade), G Protein-Coupled Receptor 61 Antibody
GPR61 (IHC-grade), G Protein-Coupled Receptor...
The GPR61 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human GRP61. It can be used to detect total GPR61 receptors in Western blots independent of phosphorylation. The GPR61 antibody can...
$ 375.00 *
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Agonist-induced Serine368 phosphorylation of the Kisspeptin Receptor
pS368-KISS1R (phospho-Kisspeptin Receptor...
Serine368 (S368) is major phosphorylation site of the Kisspeptin Receptor (KISS1R). The pS368-KISS1R antibody detects phosphorylation in response to agonists. S368 phosphorylation is likely to be involved in efficient ligand...
$ 375.00 *
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 Agonist-induced Serine473/Serine475 phosphorylation of the β1-Adrenoceptor
pS473/pS475-β1 (phospho-β1-Adrenoceptor Antibody)
Serine473/Serine475 (S473/S475) is a major phosphorylation site of the β1 adrenoceptor. The pS473/pS475-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S473/S475...
$ 375.00 *
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Agonist-induced Serine412 phosphorylation of the β1-Adrenoceptor
pS412-β1 (phospho-β1-Adrenoceptor Antibody)
Serine412 (S412) is a major phosphorylation site of the β1 adrenoceptor. The pS412-β1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S412 phosphorylation is primarily...
$ 375.00 *
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Agonist-induced Threonine314/Threonine315 phosphorylation of GPR55
pT314/pT315-GPR55 (phospho-GPR55 Antibody)
Threonine314/Threonine315 (T314/T315) is major phosphorylation site of human GPR55 receptor. The pT314/pT315-GPR55 antibody detects phosphorylation in response to agonists. T314/T315) phosphorylation is likely to be involved in efficient...
$ 375.00 *
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Agonist-induced Serine348/Threonine349 phosphorylation of the Angiotensin Receptor 1
pS348/pT349-AT1 (phospho-Angiotensin Receptor 1...
Serine348/Threonine349 (S348/T349) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS348/pT349-AT1 antibody detects phosphorylation in response to agonists. S348/T349 phosphorylation is likely to be involved in...
$ 375.00 *
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Agonist-induced Serine326/Serine328 phosphorylation of the Angiotensin Receptor 1
pS326/pS328-AT1 (phospho-Angiotensin Receptor 1...
Serine326/Serine328 (S328/S328) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS326/pS328-AT1 antibody detects phosphorylation in response to agonists. S328/S328 phosphorylation is likely to be involved in...
$ 375.00 *
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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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