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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 mouse S1P5 Receptor in transfected HEK293 cells
mS1P5 (GP-non-phospho), Mouse Sphingosine...
The non-phospho-mS1P5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouseS1P5. It can be used to detect total mS1P5 receptors in Western blots independent of phosphorylation. The mS1P5 antibody can...
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NEW
Agonist-induced Threonine315 phosphorylation of the GPR139 Receptor
pT315-GPR139 (phospho-GPR139 Antibody)
Threonine315 (T315) is major phosphorylation site of the GPR139 Receptor (GPR139). The pT315-GPR139 antibody detects phosphorylation in response to agonists. T315 phosphorylation is likely to be involved in efficient ligand sequestration.
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Immunohistochemical identification of MRGPRX3 Receptor in duodenum
MRGPRX3 (IHC-grade), MRGPRX3 Mas-related...
The non-phospho-MRGPRX3 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human MRGPRX3. It can be used to detect total MRGPRX3 receptors in Western blots independent of phosphorylation. The MRGPRX3...
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Immunohistochemical identification of Tachykinin Receptor 2 in duodenum
NK2 (IHC-grade), Tachykinin Receptor 2 Antibody
The NK2 antibody is directed against the distal end of the carboxyl-terminal tail of human Tachykinin Receptor 2. It detects selectively the NK2 Tachykinin Receptor. It can be used to detect total NK2 receptors in Western blots...
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NEW
Validation of the EP3 Prostanoid Receptor in transfected HEK293 cells
EP3 (non-phospho), EP3 Prostanoid Receptor...
The non-phospho-EP3 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human EP3. It can be used to detect total EP3 receptors in Western blots independent of phosphorylation.The EP3 antibody can also...
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Validation of the XCR Chemokine Receptor 1 in transfected HEK293 cells
XCR1 (non-phospho), XCR Chemokine Receptor 1...
The non-phospho-XCR1 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human XCR1. It can be used to detect total XCR1 receptors in Western blots independent of phosphorylation. The non-phospho-XCR1...
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Validation of the Neuropeptide FF Receptor 1 in transfected HEK293 cells
NPFF1 (non-phospho), Neuropeptide FF Receptor 1...
The NPFF1 antibody is directed against the distal end of the carboxyl-terminal tail of human NPFF1. It detects selectively the NPFF1. It can be used to detect total NPFF1 receptors in Western blots independent of phosphorylation. The...
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Validation of the Tachykinin Receptor 3 in transfected HEK293 cells
NK3 (non-phospho), Tachykinin Receptor 3 Antibody
The NK3 antibody is directed against the distal end of the carboxyl-terminal tail of human Tachykinin Receptor 3. It detects selectively the NK3 Tachykinin Receptor. It can be used to detect total NK3 receptors in Western blots...
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Immunohistochemical varification of 5-Hydroxytryptamine Receptor 6 antibody in Medial Septum
5-HT6 (IHC-grade), 5-HT Receptor 6 Antibody
The 5-HT6 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human 5-Hydroxytryptamine Receptor 6 (5-HT6). It can be used to detect total 5-HT6 receptors in Western blots independent of...
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Immunohistochemical identification of Dopamine Receptor 1 in striatum
D1 (IHC-grade), Dopamine Receptor 1 Antibody
The D1 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human D1. It can be used to detect total D1 receptors in Western blots independent of phosphorylation. The D1 antibody can also be used to...
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Validation of the S1P5 Receptor in transfected HEK293 cells.
S1P5 (non-phospho), Sphingosine 1-Phosphate...
The non-phospho-S1P5 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human S1P5. It can be used to detect total S1P5 receptors in Western blots independent of phosphorylation. The S1P5 antibody can...
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Agonist-induced Serine338/Threonine342 phosphorylation of MCH1 Receptor.
pS338/pT342-MCH1 (phospho-Melanin-Concentrating...
Serine338/Threonine342 (S338/T342) is a major phosphorylation site of the MCH1 receptor. The pS338/pT342-MCH1 antibody detects phosphorylation in response to high-efficacy agonists. S338/T342 phosphorylation is a key regulator of MCH1...
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Agonist-induced Threonine348/Serine350 phosphorylation of MCH1 Receptor
pT348/pS350-MCH1 (phospho-Melanin-Concentrating...
Threonine348/Serine350 (T348/S350) is a major phosphorylation site of the MCH1 receptor. The pT348/pS350-MCH1 antibody detects phosphorylation in response to high-efficacy agonists. T348/S350 phosphorylation is a key regulator of MCH1...
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Agonist-induced Threonine335/Serine338 phosphorylation of MCH2 Receptor
pT335/pS338-MCH2 (phospho-Melanin-Concentrating...
Threonine335/Serine338 (T335/S338) is a major phosphorylation site of the MCH2 receptor. The pT335/pS338-MCH2 antibody detects phosphorylation in response to high-efficacy agonists. T335/S338 phosphorylation is a key regulator of MCH2...
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GPR139 (non-phospho), G protein-coupled Receptor 139 Antibody
GPR139 (non-phospho), G protein-coupled...
The non-phospho-GPR139 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human GPR139. It can be used to detect total GPR139 receptors in Western blots independent of phosphorylation. The GPR139...
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Detection of non-phosphorylated human GPR35 Receptor
GPR35 (non-phospho), G Protein-coupled Receptor...
The non-phospho-GPR35 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human GPR35. It can be used to detect total GPR35 receptors in Western blots independent of phosphorylation. The...
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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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