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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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Citations
Agonist-induced Threonine348 phosphorylation of the Somatostatin Receptor 3
pT348-SST3 (phospho-Somatostatin Receptor 3...
Threonine348 (T348) is a major phosphorylation site of the somatostatin receptor 3 (SST3). The pT348-SST3 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T348 phosphorylation...
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Agonist-induced Serine337/Threonine341 phosphorylation of the Somatostatin Receptor 3.
pS337/pT341-SST3 (phospho-Somatostatin Receptor...
Serine337/Threonine341 (S337/T341) is a major phosphorylation site of the somatostatin receptor 3 (SST3). The pS337/pT341-SST3 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation....
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Agonist-induced Threonine356/Threonine359 phosphorylation of the Somatostatin Receptor 2.
pT356/pT359-SST2 (phospho-Somatostatin Receptor...
Threonine356/Threonine359 (T356/T359) is a major phosphorylation site of the somatostatin receptor 2 (SST2). The pT356/pT359-SST2 antibody detects phosphorylation in response to high-efficacy agonists such as octreotide and low-efficacy...
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Agonist-induced Threonine353/Threonine354 phosphorylation of the Somatostatin Receptor 2.
pT353/pT354-SST2 (phospho-Somatostatin Receptor...
Threonine353/Threonine354 (T353/T354) is a major phosphorylation site of the somatostatin receptor 2 (SST2). The pT353/pT354-SST2 antibody detects phosphorylation in response to high-efficacy agonists such as octreotide and low-efficacy...
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Agonist-induced Serine341/Serine343 phosphorylation of the Somatostatin Receptor 2.
pS341/pS343-SST2 (phospho-Somatostatin Receptor...
Serine341/Serine343 (S341/S343) is a major phosphorylation site of the somatostatin receptor 2 (SST2). The pS341/pS343-SST2 antibody detects phosphorylation in response to high-efficacy agonists such as octreotide and low-efficacy...
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Validation of the Somatostatin Receptor 1 in transfected HEK293 cells
SST1 (non-phospho-Somatostatin Receptor 1...
The SST1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Somatostatin Receptor 1. It can be used to detect total SST1 receptors in Western blots independent of phosphorylation. The SST1...
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Validation of the Proteinase-Activated Receptor 1 in transfected HEK293 cells
PAR1 (non-phospho), Proteinase-Activated...
The PAR1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Proteinase-Activated Receptor 1. It can be used to detect total PAR1 receptors in Western blots independent of phosphorylation. The PAR1...
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Immunohistochemical identification of Neurotensin Receptor 2 in gastric mucosa
NTS2 (IHC-grade), Neurotensin Receptor 2 Antibody
The NTS2 antibody is directed against the distal end of the carboxyl-terminal tail of human Neurotensin Receptor 2. It can be used to detect total NTS2 receptors in Western blots independent of phosphorylation. The NTS2 antibody can also...
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Validation of the Endothelin Receptor A in transfected HEK293 cells.
ETA (non-phospho-Endothelin Receptor A Antibody)
The ETA receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human endothelin receptor A. It can be used to detect ETA receptors in Western blots in a phosphorylation-sensitive manner...
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Agonist-induced Serine350/Threonine352 phosphorylation of the Atypical Chemokine Receptor 3/CXC Chemokine Receptor 7
pS350/pT352-ACKR3 (phospho-Atypical Chemokine...
Serine350/Threonine352 (S350/T352) is major phosphorylation site of the Atypical Chemokine Receptor 3 (ACKR3, previously called CXCR7). The pS350/pT352-ACKR3 antibody detects phosphorylation in response to agonists and after PKC...
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Agonist-induced Serine437/Threonine439 phosphorylation of CCK2 Receptor.
pS437/pT439-CCK2 (phospho-Cholecystokinin...
Serine437/Threonine439 (S437/T439) is a major phosphorylation site of the CCK2 receptor. The pS437/pT439-CCK2 antibody detects phosphorylation in response to high-efficacy agonists. S437/T439 phosphorylation is a key regulator of CCK2...
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Agonist-induced and Agonist-independent Threonine427/Serine429 phosphorylation of CCK2 Receptor
pT427/pS429-CCK2 (phospho-Cholecystokinin...
Threonine427/Serine429 (T427/S429) is a major phosphorylation site of the CCK2 receptor. The pT427/pS429-CCK2 antibody detects phosphorylation in response to high-efficacy agonists. T427/S429 phosphorylation is a key regulator of CRF1...
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Agonist-induced Serine417/Serine419 phosphorylation of CCK1 Receptor
pS417/pS419-CCK1 (phospho-Cholecystokinin...
Serine417/Serine419 (S417/S419) is a major phosphorylation site of the CCK1 receptor. The pS417/pS419-CCK1 antibody detects phosphorylation in response to high-efficacy agonists. S417/S419 phosphorylation is a key regulator of CCK1...
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Agonist-induced Serine412/Serine414 phosphorylation of CCK1 Receptor
pS412/pS414-CCK1 (phospho-Cholecystokinin...
Serine412/Serine414 (S412/S414) is a major phosphorylation site of the CCK1 receptor. The pS412/pS414-CCK1 antibody detects phosphorylation in response to high-efficacy agonists. S412/S414 phosphorylation is a key regulator of CCK1...
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Agonist-induced Threonine408/Threonine409 phosphorylation of CCK1 Receptor
pT408/pT409-CCK1 (phospho-Cholecystokinin...
Threonine408/Threonine409 (T408/T409) is a major phosphorylation site of the CCK1 receptor. The pT408/pT409-CCK1 antibody detects phosphorylation in response to high-efficacy agonists. T408/T409 phosphorylation is a key regulator of CCK1...
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Validation of the Chemokine Receptor CX3CR1 in transfected HEK293 cells.
CX3CR1 (non-phospho), CX3C Chemokine Receptor 1...
The non-phospho-CX3CR1 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human CX3CR1. It can be used to detect total CX3CR1 receptors in Western blots independent of phosphorylation.
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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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