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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 Threonine349/Serine350 phosphorylation of the FFA Receptor 4
pT349/pS350-FFA4 (phospho-FFA4 Antibody)
Threonine349/Serine350 (T349/S350) is major phosphorylation site of the FFA Receptor 4 (FFA4). The pT349/pS350-FFA4 antibody detects phosphorylation in response to agonists. T349/S350 phosphorylation is likely to be involved in efficient...
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NEW
Agonist-induced Threonine372/Serine373 phosphorylation of the Bradykinin Receptor 2
pT372/pS373-B2 (phospho-Bradykinin Receptor 2...
Threonine372/Serine373 (T372/S373) is major phosphorylation site of the Bradykinin Receptor 2 (B2). The pT372/pS373-B2 antibody detects phosphorylation in response to agonists. T372/S373 phosphorylation is likely to be involved in...
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Agonist-induced Serine366/Threonine369 phosphorylation of the Bradykinin Receptor 2
pS366/pT369-B2 (phospho-Bradykinin Receptor 2...
Serine366/Threonine369 (S366/T369) is major phosphorylation site of the Bradykinin Receptor 2 (B2). The pS366/pT369-B2 antibody detects phosphorylation in response to agonists. S366/T369 phosphorylation is likely to be involved in...
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Agonist-induced Threonine359/Serine360 phosphorylation of the Bombesin Receptor 2
pT359/pS360-BB2 (phospho-Bombesin Receptor 2...
Threonine359/Serine360 (T359/S360) is major phosphorylation site of the Bombesin Receptor 2 (BB2). The pT359/pS360-BB2 antibody detects phosphorylation in response to agonists. T359/S360 phosphorylation is likely to be involved in...
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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 Serine450/Serine452 phosphorylation of the PACAP Receptor 1
pS450/pS452-PAC1 (phospho-PACAP Receptor 1...
Serine450/Serine452 (S450/S452) is major phosphorylation site of the PACAP Receptor 1 (PAC1). The pS450/pS452-PAC1 antibody detects phosphorylation in response to agonists. S450/S452 phosphorylation is likely to be involved in efficient...
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Agonist-induced Threonine444/Serine447 phosphorylation of the PACAP Receptor 1
pT444/pS447-PAC1 (phospho-PACAP Receptor 1...
Threonine444/Serine447 (T444/S447) is major phosphorylation site of the PACAP Receptor 1 (PAC1). The pT444/pS447-PAC1 antibody detects phosphorylation in response to agonists. T444/S447 phosphorylation is likely to be involved in...
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Agonist-induced Serine437/Serine438 phosphorylation of the PACAP Receptor 1
pS437/pS438-PAC1 (phospho-PACAP Receptor 1...
Serine437/Serine438 (S437/S438) is major phosphorylation site of the PACAP Receptor 1 (PAC1). The pS437/pS438-PAC1 antibody detects phosphorylation in response to agonists. S437/S438 phosphorylation is likely to be involved in efficient...
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Agonist-induced Serine347/Serine348 phosphorylation of the Apelin Receptor
pS347/pS348-APLNR (phospho-Apelin Receptor...
Serine347/Serine348 (S347/S348) is major phosphorylation site of the Apelin Receptor (APLNR). The pS347/pS348-APLNR antibody detects phosphorylation in response to agonists. S347/S348 phosphorylation is likely to be involved in efficient...
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Agonist-induced Threonine353 phosphorylation of the Chemokine Receptor 10
pT353-CCR10 (phospho-Chemokine Receptor 10...
Threonine353 (T353) is a major phosphorylation site of the CCR10 receptor. The pT353-CCR10 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T353 phosphorylation is a key regulator...
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pS372/pS373-D1 (phospho-Dopamine Receptor 1 Antibody)
pS372/pS373-D1 (phospho-Dopamine Receptor 1...
Serine372/Serine373 (S372/S373) is a major phosphorylation site of the D1 receptor. The pS372/pS373-D1 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S372/S373...
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pT354-D1 (phospho-Dopamine Receptor 1 Antibody)
pT354-D1 (phospho-Dopamine Receptor 1 Antibody)
Threonine354 (T354) is a major phosphorylation site of the D1 receptor. The pT354-D1 antibody detects phosphorylation in response to high-efficacy agonists but not after PKC activation. T354 phosphorylation is a key regulator of D1...
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Validation of the Complement C5a Receptor 1 in transfected HEK293 cells.
C5a1 (non-phospho), Complement C5a Receptor 1...
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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Validation of the Dopamine Receptor 2 in transfected HEK293 cells.
D2 (non-phospho), Dopamine Receptor 2 Antibody
The non-phospho-D2 receptor antibody is directed against the third intracellular loop of mouse, rat and human D2 dopamine receptor. It detects both the long and short form of D2. It can be used to detect total D2 receptors in Western...
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pS317/pT318-D2 (phospho-Dopamine Receptor 2 Antibody)
pS317/pT318-D2 (phospho-Dopamine Receptor 2...
Serine317/threonine318 (S317/T318) is a major phosphorylation site of the D2 dopamine receptor. The pS317/pT318-D2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S317/T318...
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Agonist-induced Serine343/Serine345 phosphorylation of the Apelin Receptor
pS343/pS345-APLNR (phospho-Apelin Receptor...
Serine343/Serine345 (S343/S345) is major phosphorylation site of the Apelin Receptor (APLNR). The pS343/pS345-APLNR antibody detects phosphorylation in response to agonists. S343/S345 phosphorylation is likely to be involved in efficient...
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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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