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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 Serine362/Serine363/Serine364 phosphorylation of the mouse Sphingosine 1-Phosphate Receptor 5
pS362/pS363/pS364-mS1P5...
Serine362/Serine363/Serine364 (S362/S363/S364) is major phosphorylation site of the mouse Sphingosine 1-Phosphate Receptor 5 (mS1P5). The pS362/pS363/pS364-mS1P5 antibody detects phosphorylation in response to agonists. S362/S363/S364...
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Agonist-induced Serine347 phosphorylation of the mouse Sphingosine 1-Phosphate Receptor 5
pS347-mS1P5 (phospho-mouse-Sphingosine...
Serine347 (S347) is major phosphorylation site of the mouse Sphingosine 1-Phosphate Receptor 5 (S1P5). The pS347-mS1P5 antibody detects phosphorylation in response to agonists. S347 phosphorylation is likely to be involved in efficient...
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Validation of the Melanin-Concentrating Hormone Receptor 2 in transfected HEK293 cells
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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Immunohistochemical identification of Vasopressin Receptor 1B in kidney
V1B (IHC-grade), Vasopressin Receptor 1B Antibody
The non-phospho-V1B Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1B. It can be used to detect total V1B receptors in Western blots independent of phosphorylation. The non-phospho-V1B...
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Validation of the Vasopressin Receptor 1B in transfected HEK293 cells.
V1B (non-phospho), Vasopressin Receptor 1B...
The non-phospho-V1B Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1B. It can be used to detect total V1B receptors in Western blots independent of phosphorylation. The non-phospho-V1B...
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Immunohistochemical identification of Vasopressin Receptor 1A in pancreas
V1A (IHC-grade), Vasopressin Receptor 1A Antibody
The non-phospho-V1A Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1A. It can be used to detect total V1A receptors in Western blots independent of phosphorylation. The non-phospho-V1A...
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Validation of the Vasopressin Receptor 1A in transfected HEK293 cells
V1A (non-phospho), Vasopressin Receptor 1A...
The non-phospho-V1A Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human V1A. It can be used to detect total V1A receptors in Western blots independent of phosphorylation. The non-phospho-V1A...
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Validation of the Motilin Receptor in transfected HEK293 cells
MLNR (non-phospho) Motilin Receptor Antibody
The non-phospho MLNR antibody is directed against the distal end of the carboxyl-terminal tail human MLNR receptor. It can be used to detect total MLNR receptors in Western blots independent of phosphorylation. It can also be used to...
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Immunohistochemical identification of Calcium-sensing Receptor in pancreas
CaS (IHC-grade), Calcium-sensing Receptor Antibody
The non-phospho-CaS Calcium-sensing Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human CaS. It can be used to detect total CaS receptors in Western blots independent of phosphorylation. The...
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Validation of the Calcium-sensing Receptor in transfected HEK293 cells
CaS (non-phospho), Calcium-sensing Receptor...
The non-phospho-CaS Calcium-sensing Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human CaS. It can be used to detect total CaS receptors in Western blots independent of phosphorylation. The...
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Immunohistochemical identification of P2Y Purinoceptor 4 in pancreas
P2Y4 (IHC-grade), P2Y Purinoceptor 4 Antibody
The non-phospho-P2Y4 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human P2Y4. It can be used to detect total P2Y4 receptors in Western blots independent of phosphorylation. The non-phospho-P2Y4...
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Validation of the P2Y Purinoceptor 4 in transfected HEK293 cells
P2Y4 (non-phospho), P2Y Purinoceptor 4 Antibody
The non-phospho-P2Y4 Receptor Antibody is directed against the distal end of the carboxyl-terminal tail of human P2Y4. It can be used to detect total P2Y4 receptors in Western blots independent of phosphorylation. The non-phospho-P2Y4...
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Immunohistochemical identification of Calcitonin Receptor-Like Receptor in mouse brain
CALCR (IHC-grade) Calcitonin Receptor-Like...
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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Validation of the Calcitonin Receptor-Like Receptor in transfected HEK293 cells
CALCR (non-phospho) Calcitonin Receptor-Like...
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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Validation of the Calcitonin Receptor-Like Receptor in transfected HEK293 cells
CALCR (GP-non-phospho) Calcitonin Receptor-Like...
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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Validation of the Cannabinoid Receptor 2 in transfected HEK293 cells
CB2 (non-phospho) Cannabinoid Receptor 2 Antibody
The non-phospho-CB2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human CB2. It can be used to detect total CB2 receptors in Western blots independent of phosphorylation. The CB2 antibody can also...
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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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