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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 Threonine336 phosphorylation of the Complement C5a Receptor 1
pT336-C5a1 (phospho-Complement C5a Receptor 1...
Threonine336 (T336) is a major phosphorylation site of the C5a1 receptor. The pT336-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T336 phosphorylation is a key...
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Agonist-induced Threonine324/Serine327 phosphorylation of the Complement C5a Receptor 1
pS338/pT339-C5a1 (phospho-Complement C5a...
Serine338/Threonine339 (S338/T339) is a major phosphorylation site of the C5a1 receptor. The pS338/pT339-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S338/T339...
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Agonist-induced Threonine342 phosphorylation of the Complement C5a Receptor 1
pT342-C5a1 (phospho-Complement C5a Receptor 1...
Threonine342 (T342) is a major phosphorylation site of the C5a1 receptor. The pT342-C5a1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T342 phosphorylation is a key...
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KO-Validated
Immunohistochemical identification of Complement C5a Receptor 1 in human spleen.
mC5a1 (IHC-grade), Complement C5a Receptor 1...
The mouse C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse and rat C5a1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The mouse C5a1...
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Validation of the Complement C5a Receptor 1 in transfected HEK293 cells
C5a1 (GP-non-phospho), Complement C5a Receptor...
The non-phospho-C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human C5a1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The non-phospho-C5a1...
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Agonist-induced Serine338/Serine342 phosphorylation of the Angiotensin Receptor 1
pS338/pS342-AT1 (phospho-Angiotensin Receptor 1...
Serine338/Serine342 (S338/S342) is major phosphorylation site of the Angiotensin Receptor 1 (AT1). The pS338/pS342-AT1 antibody detects phosphorylation in response to agonists. S338/S342 phosphorylation is likely to be involved in...
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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...
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Validation of the Complement C5a Receptor 2 in transfected HEK293 cells.
C5a2 (non-phospho), Complement C5a Receptor 2...
The non-phospho-C5a2 receptor antibody is directed against the distal part of the carboxyl-terminal tail of human C5a2. It can be used to detect total C5a2 receptors in Western blots independent of phosphorylation. The non-phospho-C5a2...
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NEW
Immunohistochemical identification of Complement C5a Receptor 1 in human spleen.
C5a1 (IHC-grade), Complement C5a Receptor 1...
The C5a1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Complement 5a Receptor 1. It can be used to detect total C5a1 receptors in Western blots independent of phosphorylation. The C5a1...
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Agonist-induced Threonine369/Serine370 phosphorylation of the EP4 Prostanoid Receptor
pT369/pS370-EP4 (phospho-EP4 Prostanoid...
Threonine369/Serine370 (T369/S370) is major phosphorylation site of the EP4 Prostanoid Receptor (EP4). The pT369/pS370-EP4 antibody detects phosphorylation in response to agonists. T369/S370 phosphorylation is likely to be involved in...
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Agonist-induced Serine338/Serine339 phosphorylation of the Prostanoid Receptor 2.
pS338/pS339-DP2 (phospho-DP2 Prostanoid...
Serine338/Serine339 (S338/S339) is major phosphorylation site of the DP2 Prostanoid Receptor (DP2). The pS338/pS339-DP2 antibody detects phosphorylation in response to agonists. S338/S339 phosphorylation is likely to be involved in...
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Agonist-induced Serine493/Serine495 phosphorylation of the Parathyroid Hormone Receptor 1
pS493/pS495-PTH1 (phospho-Parathyroid Hormone...
Serine493/Serine495 (S493/S495) is major phosphorylation site of the Parathyroid Hormone Receptor 1 (PTH1). The pS493/pS495-PTH1 antibody detects phosphorylation in response to agonists. S493/S495 phosphorylation is likely to be involved...
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Immunohistochemical identification of Atypical Chemokine Receptor 4 in human spleen.
ACKR4 (IHC-grade), Atypical Chemokine Receptor...
The ACKR4 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Atypical Chemokine Receptor 4. It can be used to detect total ACKR4 receptors in Western blots independent of phosphorylation. The...
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Agonist-induced Serine348/Serine350 phosphorylation of the Atypical Chemokine Receptor 2
pS348/pS350-ACKR2 (phospho-Atypical Chemokine...
Serine348/Serine350 (S348/S350) is major phosphorylation site of the Atypical Chemokine Receptor 2 (ACKR2). The pS348/pS350-ACKR2 antibody detects phosphorylation in response to agonists. S348/S350 phosphorylation is likely to be...
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Agonist-induced Serine364/Serine366/Serine368 phosphorylation of the Sphingosine 1-Phosphate Receptor 4
pS364/pS366/pS368-S1P4 (phospho-Sphingosine...
Serine364/Serine366/Serine368 (S364/S366/S368) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 4 (S1P4). The pS364/pS366/pS368-S1P4 antibody detects phosphorylation in response to agonists. S364/S366/S368...
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Agonist-induced Threonine342/Serine343/Threonine345 phosphorylation of the Sphingosine 1-Phosphate Receptor 2
pT342/pS343/pT345-S1P2 (phospho-Sphingosine...
Threonine342Serine343/Threonine345 (T342/S343/T345) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 2 (S1P2). The pT342/pS343/pT345-S1P2 antibody detects phosphorylation in response to agonists. T342/S343/T345...
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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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