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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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SAMPLE PACK
Agonist-induced Serine355/Serine356 phosphorylation of the β2-Adrenoceptor
β2-Adrenoceptor Sample Pack (phospho- and...
β2-Adrenoceptor Antibody Sample Pack consisting of three phospho- and one non-phospho-β2-Adrenoceptor Antibodies 4 x 20 µL trial size each. Specifically, this sample pack contains the following antibodies pS355/pS356-β2 (7TM0029A),...
300.00 € *
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SAMPLE PACK
Agonist-induced Serine331/Threonine332 phosphorylation of the Angiotensin Receptor 1
AT1 Sample Pack (phospho- and...
AT1 Sample Pack consisting of all four available phospho- and non-phospho-AT1 Antibodies 4 x 20 µL trial size each. Specifically, this sample pack contains the following antibodies pS331/pT332-AT1 (7TM0034A), pS335/pT336-AT1 (7TM0034B),...
300.00 € *
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Agonist-induced Serine385/Serine387 phosphorylation of the Neuropeptide Receptor 1
pS385/pS387-NPFF1 (phospho-Neuropeptide FF...
Serine385/Serine387 (S385/S387) is major phosphorylation site of the Neuropeptide FF Receptor 1 (NPFF1). The pS385/pS387-NPFF1 antibody detects phosphorylation in response to agonists. S385/S387 phosphorylation is likely to be involved...
375.00 € *
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NEW
Agonist-induced Serine355/Serine356 phosphorylation of the β2 Adrenoceptor
pS355/pS356-β2 (phospho-β2-Adrenoceptor Antibody)
Serine355/Serine356 (S355/S356) is a major phosphorylation site of the β2 adrenoceptor. The pS355/pS356-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S355/S356...
375.00 € *
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NEW
Agonist-induced Serine257/Threonine262 phosphorylation of the D3 Dopamine Receptor
pS257/pT262-D3 (phospho-Dopamine Receptor 3...
Serine257/Threonine262 (S257/T262) is a major phosphorylation site of the D3 dopamine receptor. The pS257/pT262-D3 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S262/T262...
375.00 € *
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NEW
Agonist-induced Serine342 phosphorylation of the Chemokine Receptor 5
pS342-CCR5 (phospho-Chemokine Receptor 5 Antibody)
Serine342 (S342) is a major phosphorylation site of the CCR5 receptor. The pS342-CCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. S342 phosphorylation is a key regulator of...
375.00 € *
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Agonist-induced Threonine340 phosphorylation of the Chemokine Receptor 5
pT340-CCR5 (phospho-Chemokine Receptor 5 Antibody)
Threonine340 (T340) is a major phosphorylation site of the CCR5 receptor. The pT340-CCR5 antibody detects phosphorylation in response to high- and low-efficacy agonists and after PKC activation. T340 phosphorylation is a key regulator of...
375.00 € *
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NEW
Agonist-induced Serine355 phosphorylation of the β2-Adrenoceptor
pS355-β2 (phospho-β2-Adrenoceptor Antibody)
Serine355 (S355) is a major phosphorylation site of the β2 adrenoceptor. The pS355-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S355 phosphorylation is primarily...
375.00 € *
Details
NEW
Agonist-induced Serine356 phosphorylation of the β2-Adrenoceptor
pS356-β2 (phospho-β2-Adrenoceptor Antibody)
Serine356 (S356) is a major phosphorylation site of the β2 adrenoceptor. The pS356-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. S356 phosphorylation is primarily...
375.00 € *
Details
NEW
Agonist-induced Threonine360 phosphorylation of the β2-Adrenoceptor
pT360-β2 (phospho-β2-Adrenoceptor Antibody)
Threonine360 (T360) is a major phosphorylation site of the β2 adrenoceptor. The pT360-β2 antibody detects phosphorylation in response to high- and low-efficacy agonists but not after PKC activation. T360 phosphorylation is primarily...
375.00 € *
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NEW
Agonist-induced Serine351/Serine353 phosphorylation of the Sphingosine 1-Phosphate Receptor 1
pS351/pS353-S1P1 (phospho-Sphingosine...
Serine351/Serine353 (S351/S353) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 1 (S1P1). The pS351/pS353-S1P1 antibody detects phosphorylation in response to agonists. S351/S353 phosphorylation is likely to be...
375.00 € *
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NEW
Agonist-induced Serine330/Serine331 phosphorylation of the Sphingosine 1-Phosphate Receptor 2
pS330/pS331-S1P2 (phospho-Sphingosine...
Serine330/Serine331 (S330/S331) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 2 (S1P2). The pS330/pS331-S1P2 antibody detects phosphorylation in response to agonists. S330/S331 phosphorylation is likely to be...
375.00 € *
Details
NEW
Agonist-induced Serine366/Serine367 phosphorylation of the Sphingosine 1-Phosphate Receptor 5
pS366/pS367-S1P5 (phospho-Sphingosine...
Serine366/Ser367 (S366/S367) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 5 (S1P5). The pS366/pS367-S1P5 antibody detects phosphorylation in response to agonists. S366/S367 phosphorylation is likely to be...
375.00 € *
Details
NEW
Agonist-induced Serine353/Serine355 phosphorylation of the Sphingosine 1-Phosphate Receptor 1
pS353/pS355-S1P1 (phospho-Sphingosine...
Serine353/Serine355 (S353/S355) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 1 (S1P1). The pS353/pS355-S1P1 antibody detects phosphorylation in response to agonists. S353/S355 phosphorylation is likely to be...
375.00 € *
Details
NEW
Agonist-induced Serine358/Serine359 phosphorylation of the Sphingosine 1-Phosphate Receptor 1
pS358/pS359-S1P1 (phospho-Sphingosine...
Serine358/Serine359 (S358/S359) is major phosphorylation site of the Sphingosine 1-Phosphate Receptor 1 (S1P1). The pS358/pS359-S1P1 antibody detects phosphorylation in response to agonists. S358/S359 phosphorylation is likely to be...
375.00 € *
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NEW
pT342/pS343/pT345-S1P2 (phospho-Sphingosine 1-Phosphate Receptor 2 Antibody)
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...
375.00 € *
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13 From 28

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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