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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
 Validation of the Cholecystokinin Receptor 2 in transfected HEK293 cells
CCK2 (non-phospho), Cholecystokinin Receptor 2...
The CCK2 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Cholecystokinin Receptor 2 (previously called CCKB). It can be used to detect total CCK2 receptors in Western blots independent of...
$ 300.00 *
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pS417/pS419-CCK1 (phospho-Cholecystokinin Receptor 1 Antibody)
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...
$ 375.00 *
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pS412/pS414-CCK1 (phospho-Cholecystokinin Receptor 1 Antibody)
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...
$ 375.00 *
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pT408/pT409-CCK1 (phospho-Cholecystokinin Receptor 1 Antibody)
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...
$ 375.00 *
Citations
Validation of the Cholecystokinin Receptor 1 in transfected HEK293 cells
CCK1 (non-phospho), Cholecystokinin Receptor 1...
The CCK1 receptor antibody is directed against the distal end of the carboxyl-terminal tail of human Cholecystokinin Receptor 1 (previously called CCKA). It can be used to detect total CCK1 receptors in Western blots independent of...
$ 375.00 *
Citations
Agonist-induced Serine324/Serine325 phosphorylation of the CXC Chemokine Receptor 4.
pS324/pS325-CXCR4 (phospho-CXC Chemokine...
Serine324/serine325 (S324/S325) is a major phosphorylation site of the CXCR4 receptor. The pS324/pS325-CXCR4 antibody detects phosphorylation in response to agonists but not after PKC activation. S324/S325 phosphorylation is a key...
$ 375.00 *
Citations
pS346/pS347-CXCR4 (phospho-CXC Chemokine Receptor 4 Antibody)
pS346/pS347-CXCR4 (phospho-CXC Chemokine...
Serine346/serine347 (S346/S347) is a major phosphorylation site of the CXCR4 receptor. The pS346/pS347-CXCR4 antibody detects phosphorylation in response to agonists and after PKC activation. S346/S347 phosphorylation is a key regulator...
$ 375.00 *
Citations
KO-Validated
Validation of the CXC Chemokine Receptor 4 in transfected HEK293 cells.
CXCR4 (non-phospho), CXC Chemokine Receptor 4...
The non-phospho-CXCR4 receptor antibody is directed against the distal end of the carboxyl-terminal tail of mouse, rat and human CXCR4. It can be used to detect CXCR4 receptors in Western blots in a phosphorylation-sensitive manner....
$ 375.00 *
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 Agonist-induced Serine330 phosphorylation of the CXC Chemokine Receptor 4
pS330-CXCR4 (phospho-CXC Chemokine Receptor 4...
Serine330(S330) is a major phosphorylation site of the CXCR4 receptor. The pS330-CXCR4 antibody detects phosphorylation in response to agonists and after PKC activation. S330 phosphorylation is a key regulator of CXCR4 desensitization,...
$ 375.00 *
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Agonist-induced Serine338/Serine339 phosphorylation of the CXC Chemokine Receptor 4
pS338/pS339-CXCR4 (phospho-CXC Chemokine...
Serine338 and Serine339 (S338/S339) are a major phosphorylation sites of the CXCR4 receptor. The pS338/pS339-CXCR4 antibody detects phosphorylation in response to agonists and after PKC activation. S338/S339 phosphorylation is a key...
$ 375.00 *
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Validation of Atypical Chemokine Receptor 4 in transfected HEK293 cells
ACKR4 (non-phospho), Atypical Chemokine...
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...
$ 375.00 *
Citations
Validation of the Atypical Chemokine Receptor 3/CXC Chemokine Receptor 7 in transfected HEK293 cells.
ACKR3 (non-phos, C-Term), Atypical Chemokine...
The non-phospho-ACKR3 receptor antibody is directed against the carboxyl-terminal tail of mouse, rat and human ACKR3/CXCR7. It can be used to detect total ACKR3 receptors in Western blots independent of phosphorylation. The...
$ 300.00 *
Citations
pS350/pT352-ACKR3 (phospho-Atypical Chemokine Receptor 3 Antibody)
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...
$ 375.00 *
NEW
Validation of the Atypical Chemokine Receptor 3/CXC Chemokine Receptor 7 in transfected HEK293 cells
ACKR3 (non-phos, N-Term), Atypical Chemokine...
The non-phospho-ACKR3 receptor antibody is directed against the amino-terminal tail of mouse, rat and human ACKR3/CXCR7. It can be used to detect total ACKR3 receptors in Western blots independent of phosphorylation. The...
$ 300.00 *
NEW
Agonist-induced Serine355/Serine360 phosphorylation of the Atypical Chemokine Receptor 3/CXC Chemokine Receptor 7
pS355/pS360-ACKR3 (phospho-Atypical Chemokine...
Serine355/Serine360 (S355/S360) is major phosphorylation site of the Atypical Chemokine Receptor 3 (ACKR3, previously called CXCR7). The pS355/pS360-ACKR3 antibody detects phosphorylation in response to agonists. S355/S360...
$ 375.00 *
NEW
Agonist-induced Serine345/Serine346 phosphorylation of Chemerin Receptor 1
pS345/pS346-CMKLR1 (phospho-Chemerin Receptor 1...
Serine345/Serine346 (S345/S346) is a major phosphorylation site of the Chemerin receptor 1 (CMKLR1). The pS345/pS346-CMKLR1 antibody detects phosphorylation in response to high-efficacy agonists. S345/S346 phosphorylation is a key...
$ 375.00 *
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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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