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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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Agonist-induced Threonine370 phosphorylation of the Bombesin Receptor 3
pT370-BB3 (phospho-Bombesin Receptor 3 Antibody)
Threonine370 (T370) is major phosphorylation site of the Bombesin Receptor 3 (BB3). The pT370-BB3 antibody detects phosphorylation in response to agonists. T370 phosphorylation is likely to be involved in efficient ligand sequestration...
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Immunohistochemical Identification of Agonist-induced Serine425 phosphorylation of the Cannabinoid Receptor 1 in mouse brain
pS425-CB1 (IHC-grade phospho-Cannabinoid...
Serine425 (S425) is major phosphorylation site of the Cannabinoid Receptor 1 (CB1). The pS425-CB1 antibody detects phosphorylation in response to agonists. S425 phosphorylation is likely to be involved in efficient ligand sequestration...
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Immunohistochemical identification of Orexin Receptor 1 in pancreas
OX1 (IHC-grade) Orexin Receptor 1 Antibody
The non-phospho OX1 receptor antibody is directed against the distal end of the carboxyl-terminal tail human OX1 receptor. It can also be used to isolate and enrich OX1 receptors from tissue lysates. It also detects OX1 in cultured cells...
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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...
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Agonist-induced Threonine389/Threonine390 phosphorylation of the NK3 Receptor3.
pT389/pT390-NK3 (phospho-Tachykinin Receptor 3...
Threonine389/Threonine390 (T389/T390) is a major phosphorylation site of the NK3 receptor. The pT389/pT390-NK3 antibody detects phosphorylation in response to high-efficacy agonists. T389/T390 phosphorylation is a key regulator of NK3...
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Agonist-induced Serine398/Serine399 phosphorylation of the NK 3 Recept
pS398/pS399-NK3 (phospho-Tachykinin Receptor 3...
Serine398/Serine399 (S398/S399) is major phosphorylation site of the Tachykinin Receptor 3 (NK3). The pS398/pS399-NK3 antibody detects phosphorylation in response to agonists. S398/S399 phosphorylation is likely to be involved in...
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Agonist-induced Threonine402/Threonine404 phosphorylation of the NK 3 Receptor.
pT402/pT404-NK3 (phospho-Tachykinin Receptor 3...
Threonine402/Threonine404 (T402/T404) is a major phosphorylation site of the NK3 receptor. The pT402/pT404-NK3 antibody detects phosphorylation in response to high-efficacy agonists. T402/T404 phosphorylation is a key regulator of NK3...
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Immunohistochemical identification of Proteinase-Activated Receptor 2 in Kidney
PAR2 (IHC-grade), Proteinase-Activated Receptor...
The PAR2 antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human PAR2. It can be used to detect total PAR2 receptors in Western blots independent of phosphorylation. The PAR2 antibody can also be...
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Agonist-induced Serine360/Serine361 phosphorylation of the Urotensin Receptor.
pS360/pS361-UT (phospho-Urotensin Receptor...
Serine360/Serine361 (S360/S361) is major phosphorylation site of the Urotensin Receptor (UT). The pS360/pS361-UT antibody detects phosphorylation in response to agonists. S360/S361 phosphorylation is likely to be involved in efficient...
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Agonist-induced Threonine343/Serine347/Threonine348 phosphorylation of the 5-Hydroxytryptamine Receptor 4
pT343/pS347/pT348-5-HT4 (phospho-5-HT Receptor...
Threonine343/Serine347/Threonine348 is a major phosphorylation site of the 5-HT4 receptor. The pT343/pS347/pT348-5-HT4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T343/S347/T348...
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Agonist-induced Threonine311/Serine314 phosphorylation of the M4 Muscarinic Acetylcholine Receptor
pT311/pS314-M4 (phospho-M4 Muscarinic...
Threonine311/Serine314 is a major phosphorylation site of the M4 Muscarinic Acetylcholine Receptor. The pT311/pS314-M4 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T311/S314...
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Validation of the Melanin-Concentrating Hormone Receptor 1 in transfected HEK293 cells
MCH1 (non-phospho) Melanin-Concentrating...
The non-phospho MCH1 receptor antibody is directed against the distal end of the carboxyl-terminal tail human MCH1 receptor. It can be used to detect total MCH1 receptors in Western blots independent of phosphorylation. It can also be...
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Immunohistochemical identification of Melanin-Concentrating Hormone Receptor 1 in pancreas
MCH1 (IHC-grade) Melanin-Concentrating Hormone...
The non-phospho MCH1 receptor antibody is directed against the distal end of the carboxyl-terminal tail human MCH1 receptor. It can be used to detect total MCH1 receptors in Western blots independent of phosphorylation. It can also be...
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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 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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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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11 From 42

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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